논문현황

    • 2015-10-06
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    The preventive and therapeutic effects of intravenous human adipose-derived stem cells in Alzheimer’s disease mice

    Alzheimer’s disease (AD) is characterized by the accumulation of amyloid plaques and neurofibrillary tangles accompanied by cognitive dysfunction. The aim of the present study was to elucidate preventive and therapeutic potential of stem cells for AD. Among stem cells, autologous human adipose-derived stem cells (hASCs) elicit no immune rejection responses, tumorigenesis, or ethical problems. We found that intravenously transplanted hASCs passed through the BBB and migrated into the brain. The learning, memory and pathology in an AD mouse model (Tg2576) mice greatly improved for at least 4 months after intravenous injection of hASC. The number of amyloid plaques and Ab levels decreased significantly in the brains of hASC-injected Tg mice compared to those of Tg-sham mice. Here, we first report that intravenously or intracerebrally transplanted hASCs significantly rescues memory deficit and neuropathology, in the brains of Tg mice by upregulating IL-10 and VEGF and be a possible use for the prevention and treatment of AD

    • 2015-10-06
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    Intra-articular Injection of Mesenchymal Stem Cells for the Treatment of Osteoarthritis of the Knee: A Proof-of-Concept Clinical Trial

    요약 :
    Mesenchymal stem cells (MSCs) are known to have a potential for articular cartilage regeneration. However, most studies focused on focal cartilage defect through surgical implantation. For the treatment of generalized cartilage loss in osteoarthritis, an alternative delivery strategy would be more appropriate. The purpose of this study was to assess the safety and efficacy of intra-articular injection of autologous adipose tissue derived MSCs (AD-MSCs) for knee osteoarthritis. We enrolled 18 patients with osteoarthritis of the knee and injected AD MSCs into the knee. The phase I study consists of three dose-escalation cohorts; the low-dose (1.0 3 107 cells), mid-dose (5.0 3 107), and high-dose (1.0 3 108) group with three patients each. The phase II included nine patients receiving the high-dose. The primary outcomes were the safety and the Western Ontario and McMaster Universities Osteoarthritis index (WOMAC) at 6 months. Secondary outcomes included clinical, radiological, arthroscopic, and histological evaluations. There was no treatment-related adverse event.
    The WOMAC score improved at 6 months after injection in the high-dose group. The size of cartilage defect decreased while the volume of cartilage increased in the medial femoral and tibial condyles of the high-dose group. Arthroscopy showed that the size of cartilage defect decreased in the medial femoral and medial tibial condyles of the high-dose group. Histology demonstrated thick, hyaline-like cartilage regeneration. These results showed that intra-articular injection of 1.0 3 108 AD MSCs into the osteoarthritic knee improved function and pain of the knee joint without causing adverse events, and reduced cartilage defects by regeneration of hyaline-like articular cartilage

    • 2015-10-06
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    Long-term Follow-up after Implantation of Autologous Adipose Tissue Derived Mesenchymal Stem Cells to Treat a Dog with Stifle Joint Osteoarthrosis

    요약 : 
    A 5-year-old castrated male Chihuahua weighing 1.54 kg was examined because of a several month history of progressive right hind limb lameness. Physical examination of the stifle joints revealed pain and a grade IV medial patellar luxation on the right stifle joint. The right and left stifle joints were associated with a lameness of grade 2 and grade 0, respectively. Radiography revealed osteophytes or subchondral cystic lesions on the right and left stifle joints. Osteoarthrosis (OA) scores for the right and left stifle joints were 20 and 12 respectively. Combination of surgery and implantation of autologous adipose tissue derived mesenchymal stem cells (aAT-MSCs) was determined with informed consent. 1 × 106 aAT-MSCs suspended in PBS and 0.6 mL of hyaluronic acid were injected in the right stifle joint postoperatively. Osteoarthrosis scores and the lameness grade for the right and left stifle joints were 19 and 13, and 0 and 0 19 months after treatment, respectively, and 14 and 15, and 0 and 0 five years after treatment, respectively. This case report shows radiographical evidence of a decrease in osteophytes and subchondral cystic lesions on the stifle joint with OA after aAT-MSCs injection.

    • 2015-10-06
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    Establishment of efficacy and safety assessment of human adipose tissue-derived mesenchymal stem cells (hATMSCs) in a nude rat femoral segmental defect model

    요약 : 
    Human adipose tissue-derived mesenchymal stem cell (hATMSC) have emerged as a potentially powerful tool for bone repair, but an appropriate evaluation system has not been established. The purpose of this study was to establish a preclinical assessment system to evaluate the efficacy and safety of cell therapies in a nude rat bone defect model. Segmental defects (5 mm) were created in the femoral diaphyses and transplanted with
    cell media (control), hydroxyapatite/tricalcium phosphate scaffolds (HA/TCP, Group I), hATMSCs (Group II), or three cell-loading density of hATMSC-loaded HA/TCP (Group III-V). Healing response was evaluated by serial radiography, micro-computed tomography and histology at 16 weeks. To address safety-concerns, we conducted a GLP-compliant toxicity study. Scanning electron microscopy studies showed that hATMSCs filled the pores/surfaces of scaffolds in a cell-loading density-dependent manner. We detected significant increases in bone formation in the hATMSC-loaded HA/TCP groups compared with other groups. The amount of new bone formation increased with increases in loaded cell number. In a toxicity study, no significant hATMSC-related changes were found in body weights, clinical signs, hematological/biochemical values, organ weights, or histopathological findings. In conclusion, hATMSCs loaded on HA/TCP enhance the repair of bone defects and was found to be safe under our preclinical efficacy/safety hybrid assessment system.

    • 2015-10-06
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    Administering human adipose-derived mesenchymal stem cells to prevent and treat experimental arthritis.

    요약 : 
    Rheumatoid arthritis is a chronic autoimmune disease and affecting approximately 1% of the population. Human adipose-derived mesenchymal stem cells (hASCs) were recently found to suppress effector T cell and inflammatory responses and, thus, to have beneficial effects in various autoimmune diseases. In this study, we examined whether hASCs could play a protective and/or therapeutic role in collagen-induced arthritis (CIA). We showed that hASCs both prevented and treated CIA by significantly reducing the incidence and severity of experimental arthritis. We further demonstrated that treatment with hASCs inhibited the production of various inflammatory mediators, decreased antigen-specific Th1/Th17 cell expansion, and induced the production of anti-inflammatory cytokine interleukin-10. Moreover, hASCs could induce the generation of antigen-specific Treg cells with the capacity to suppress collagen-specific T cell responses.

    • 2015-10-06
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    Characteristics of mouse adipose tissue-derived stem cells and therapeutic comparison between syngeneic and allogeneic adipose tissue-derived stem cell transplantation in experimental autoimmune thyroiditis

    요약 : 
    Previously, we found that the intravenous administration of human adipose tissue-derived mesenchymal stem cells was a promising therapeutic option for autoimmune thyroiditis even when the cells were transplanted into a xenogeneic model without an immunosuppressant. Therefore, we explored the comparison between the therapeutic effects of syngeneic and allogeneic adipose tissue-derived stem cells on an experimental autoimmune thyroiditis mouse model. Experimental autoimmune thyroiditis was induced in C57BL/6 mice by immunization with porcine thyroglobulin. Adipose tissue-derived stem cells derived from C57BL/6 mice (syngeneic)
    or BALB/c mice (allogeneic) or saline as a vehicle control were administered intravenously four times weekly. Blood and tissue samples were collected 1 week after the last transplantation. Adipose tissue-derived stem cells from mice were able to differentiate into multiple lineages in vitro; however, mouse adipose tissuederived stem cells did not have immunophenotypes identical to those from humans. Syngeneic and allogeneic administrations of adipose tissue-derived stem cells reduced thyroglobulin autoantibodies and the inflammatory immune response, protected against lymphocyte infiltration into the thyroid, and restored the Th1/Th2 balance without any adverse effects. However, different humoral immune responses were observed for infused cells from different stem cell sources. The strongest humoral immune response was induced by xenogeneic transplantation, followed by allogeneic and syngeneic administration, in that order. The stem cells were mostly found in the spleen, not the thyroid. This migration might be because the stem cells primarily function in systemic immune modulation, due to being given prior to disease induction. In this study, we confirmed that there were equal effects of adipose tissue-derived stem cells in treating autoimmune thyroiditis between syngeneic and allogeneic transplantations

    • 2015-10-06
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    Stem cell treatment for patients with autoimmune disease by systemic infusion of culture-expanded autologous adipose tissue derived mesenchymal stem cells.

    요약 : 
    Prolonged life expectancy, life style and environmental changes have caused a changing disease pattern in developed countries towards an increase of degenerative and autoimmune diseases. Stem cells have become a promising tool for their treatment by promoting tissue repair and protection from immune-attack associated damage. Patient-derived autologous stem cells present a safe option for this treatment since these will not induce immune rejection and thus multiple treatments are possible without any risk for allogenic sensitization, which may arise from allogenic stem cell transplantations. Here we report the outcome of treatments with culture expanded human adipose-derived mesenchymal stem cells (hAdMSCs) of 10 patients with autoimmune associated tissue damage and exhausted therapeutic options, including autoimmune hearing loss, multiple sclerosis, polymyotitis, atopic dermatitis and rheumatoid arthritis. For treatment, we developed a standardized culture-expansion protocol for hAdMSCs from minimal amounts of fat tissue, providing sufficient number of cells for repetitive injections. High expansion efficiencies were routinely achieved from autoimmune patients and from elderly donors without measurable loss in safety profile, genetic stability, vitality and differentiation potency, migration and homing characteristics. Although the conclusions that can be drawn from the compassionate use treatments in terms of therapeutic efficacy are only preliminary, the data provide convincing evidence for safety and therapeutic properties of systemically administered AdMSC in human patients with no other treatment options. The authors believe that exvivo-expanded autologous AdMSCs provide a promising alternative for treating autoimmune diseases. Further clinical studies are needed that take into account the results obtained from case studies as those presented here

    • 2017-05-08
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    Anti-inflammatory Effects of Oct4/Sox2-overexpressing Human Adipose Tissue-derived Mesenchymal Stem Cells

    Background/Aim: The transcription factors Oct4
    and Sox2 enhance the proliferation and pluripotency of
    human adipose tissue-derived mesenchymal stem cells (hATMSCs);
    however, the anti-inflammatory effects of Oct4- and
    Sox2-overexpressing hAT-MSCs (Oct4/Sox2-hAT-MSCs) are
    unclear. Here, we evaluated the anti-inflammatory effects of
    Oct4/Sox2-hAT-MSCs in vitro and in vivo.

    Accordingly, in this study, the anti-inflammatory effects
    of lentivirus-transduced Oct4/Sox2-hAT-MSCs were
    investigated by determining the expression of
    inflammation-related cytokines in macrophage cell lines
    treated with conditioned medium in vitro. In addition,
    sickness scores (diarrhea, eye condition, activity and fur
    condition) and survival rates were used to evaluate the antiinflammatory
    effects of the engineered hAT-MSCs in a
    mouse model.

    • 2015-10-06
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    Immunomodulatory effects of human amniotic membrane-derived mesenchymal stem cells

    요약 : 
    Human amniotic membrane-derived mesenchymal stem cells (hAM-MSCs) are capable of differentiating into several lineages and possess immunomodulatory properties. In this study, we investigated the soluble factor-mediated immunomodulatory effects of hAM-MSCs. Mitogen-induced peripheral blood mononuclear cell (PBMC) proliferation was suppressed by hAM-MSCs in a dose-dependent manner as well as hAM-MSC culture supernatant. Moreover, interferon-gamma and interleukin (IL)-17 production significantly decreased from PBMC, whereas IL-10 from PBMCs and transforming growth factor beta (TGF-β) production from hAM-MSCs significantly increased in co-cultures of hAM-MSCs and PBMCs. Production of several MSC factors, including hepatocyte growth factor (HGF), TGF-β, prostaglandin E2 (PGE2), and indoleamine 2, 3 dioxygenase (IDO), increased significantly in hAM-MSCs co-cultured with PBMCs. These results indicate that the immunomodulatory effects of hAM-MSCs may be associated with soluble factors (TGF-β, HGF, PGE2, and IDO), suggesting that hAM-MSCs may have potential clinical use in regenerative medicine

    • 2017-02-09
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    Mesenchymal stem cell transplantation can restore lupus disease-associated miRNA expression and Th1/Th2 ratios in a murine model of SLE

    Treatment with
    cyclophosphamide or ASC can change miRNAs and decrease miR-96-5p and miR-182-5p expression,
    as well as decreasing the CD138 proportion and the Th1/Th2 ratio, which might be involved in the
    therapeutic mechanism.

    • 2015-10-06
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    Reversal of serologic, immunologic, and histologic dysfunction in mice with systemic lupus erythematosus by long-term serial adipose tissue-derived mesenchymal stem cell transplantation.

    요약 : 
    Objective. To investigate the efficacy of human adipose tissue–derived mesenchymal stem cell (ADMSC) transplantation in systemic lupus erythematosus (SLE) and to determine the optimal transplantation
    window for stem cells either before or after disease onset.
    Methods. (NZB NZW)F1 mice with SLE were administered human AD-MSCs (5 105) intravenously every 2 weeks from age 6 weeks until age 60 weeks, while the control group received saline vehicle on the same schedule. Another experiment was carried out with a different initiation time point for serial transplantation (age 6 weeks or age 32 weeks).
    Results. Long-term serial administration (total of 28 times) of human AD-MSCs ameliorated SLE without any adverse effects. Compared with the control group, the human AD-MSC–treated group had a significantly
    higher survival rate with improvement of histologic and serologic abnormalities and immunologic function, and also had a decreased incidence of proteinuria. Anti–double-stranded DNA antibodies and blood urea nitrogen levels decreased significantly with transplantation of human AD-MSCs, and serum levels of granulocyte–macrophage colony-stimulating factor, interleukin-4 (IL-4), and IL-10 increased significantly. A significant
    increase in the proportion of CD4 FoxP3 cells and a marked restoration of capacity for cytokine production were observed in spleens from the human AD-MSC–treated group. In the second experiment, an early stage
    treatment group showed better results (higher survival rates and lower incidence of proteinuria) than an advanced stage treatment group.
    Conclusion. Serial human AD-MSC transplantation had beneficial effects in the treatment of SLE, without adverse effects. Transplantation of human ADMSCs before disease onset was preferable for amelioration of SLE and restoration of immune homeostasis.

    • 2015-10-06
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    Transplantation of Adipose Tissue-Derived Mesenchymal Stem Cells Prevents the Development of Lupus Dermatitis

    요약 : 
    MRL/lpr mice spontaneously develop high titers of anti-dsDNA antibodies and symptoms such as glomerular nephritis and organ weight gain. They also develop spontaneous skin inflammation similar to the cutaneous
    lesions common in human lupus erythematosus. This study aimed to compare the effects of long-term serial administration of human adipose tissue-derived mesenchymal stem cells (ASCs), CTLA4Ig-overexpressing
    ASCs, and cyclophosphamide treatment in MRL/lpr mice. MRL/lpr mice were divided into saline (C), cyclophosphamide (Y), ASC early (E), ASC late (L), and CTLA4Ig-overexpressing ASC (CT) treatment groups.
    Background-matched control MRL/MPJ mice treated with saline (N) were also compared. The treatment period was 5–23 weeks, except for the L group (15–23 weeks). Blood and tissue samples were collected when the mice were 24 weeks old. Organ weight, anti-dsDNA antibodies, urine protein, skin and kidney histologic abnormalities, and trabecular bone volume were evaluated. The Y group showed the greatest decrease in anti-dsDNA antibodies, organ weight, degree of kidney inflammation and glomerular infiltration of C3, and incidence rate of severe proteinuria; the E, L, and CT treatment groups showed better results than the C group. ASC transplantation reduced anti-dsDNA antibody levels significantly. Mice treated with ASCs or CTLA4Ig-ASCs starting from the early disease stage did not show dermatitis upon gross examination; they demonstrated significant improvement in hyperkeratosis, acanthosis, and inflammatory cell infiltration scores in histopathology. Micro-CT analysis revealed that cyclophosphamide treatment significantly decreased bone volume and increased bone spacing in the trabecular bone. Thus, we found that ASC and CTLA4-ASC treatments prevent lupus dermatitis development in MRL/lpr mice without adverse effects

    • 2015-10-06
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    Reduction of Liver Fibrosis by Xenogeneic Human Umbilical Cord Blood and Adipose Tissue-derived Multipotent Stem Cells without Treatment of an Immunosuppressant

    요약 : 
    Therapy using stem cells for the liver fibrosis is a prospective alternative to overcome the insufficiency of transplantable liver donor. Here, we demonstrated xenogeneic human cell therapy for the treatment of rat liver
    fibrosis without the use of an immunosuppressant. Liver fibrosis was induced by dimethylnitrosamine for 5 weeks in six-week-old male Sprague-Dawley rats. Human umbilical cord blood- and adipose tissue-derived multipotent stem cells were injected intravenously by the tail vein after one week. Blood samples were collected and liver samples were stained with Masson’s trichrome in order to evaluate the amount of fibrosis. After the cell injection, the level of total protein, albumin, alanine transaminase and aspartic acid transaminase was recovered to the similar level of the normal rats. The liver weight per body weight increased after the cell injection. Collagen fiber, near the portal triad and marginal region, was reduced, significantly. Taken together, it is suggested that xenotransplantation of multipotent stem cells might be a candidate for the treatment of liver fibrosis without the use of an immunosuppresant

    • 2015-10-06
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    The therapeutic efficacy of human adipose tissue-derived mesenchymal stem cells on experimental autoimmune hearing loss in mice.

    요약 : 
    Autoimmune inner ear disease is characterized by progressive, bilateral although asymmetric, sensorineural hearing loss. Patients with autoimmune inner ear disease had higher frequencies of interferon-c-producing
    T cells than did control subjects tested. Human adipose-derived mesenchymal stem cells (hASCs) were recently found to suppress effector T cells and inflammatory responses and therefore have beneficial effects in various autoimmune diseases. The aim of this study was to examine the immunosuppressive activity of hASCs on autoreactive T cells from the experimental autoimmune hearing loss (EAHL) murine model. Female BALB/c mice underwent b-tubulin immunization to develop EAHL; mice with EAHL were given hASCs or PBS intraperitoneally once a week for 6 consecutive weeks. Auditory brainstem responses were examined over time. The T helper type 1 (Th1)/Th17-mediated autoreactive responses were examined by determining the proliferative response and cytokine profile of splenocytes stimulated with b-tubulin. The frequency of regulatory T (Treg) cells and their suppressive capacity on autoreactive T cells were also determined. Systemic infusion of hASCs significantly improved hearing function and protected hair cells in established EAHL. The hASCs decreased the proliferation of antigen-specific Th1/Th17 cells and induced the production of anti-inflammatory cytokine interleukin-10 in splenocytes.
    They also induced the generation of antigen-specific CD4+ CD25+ Foxp3+ Treg cells with the capacity to suppress autoantigen-specific T-cell responses. The experiment demonstrated that hASCs are one of the important regulators of immune tolerance with the capacity to suppress effector T cells and to induce the generation of antigen-specific Treg cells.

    • 2015-10-06
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    Evaluation of the potential use of adipose-derived mesenchymal stromal cells in the treatment of canine atopic dermatitis: a pilot study.

    요약 : 
    Human amniotic membrane-derived mesenchymal stem cells (hAM-MSCs) are capable of differentiating into several lineages and possess immunomodulatory properties. In this study, we investigated the soluble factor-mediated immunomodulatory effects of hAM-MSCs. Mitogen-induced peripheral blood mononuclear cell (PBMC) proliferation was suppressed by hAM-MSCs in a dose-dependent manner as well as hAM-MSC culture supernatant. Moreover, interferon-gamma and interleukin (IL)-17 production significantly decreased from PBMC, whereas IL-10 from PBMCs and transforming growth factor beta (TGF-β) production from hAM-MSCs significantly increased in co-cultures of hAM-MSCs and PBMCs. Production of several MSC factors, including hepatocyte growth factor (HGF), TGF-β, prostaglandin E2 (PGE2), and indoleamine 2, 3 dioxygenase (IDO), increased significantly in hAM-MSCs co-cultured with PBMCs. These results indicate that the immunomodulatory effects of hAM-MSCs may be associated with soluble factors (TGF-β, HGF, PGE2, and IDO), suggesting that hAM-MSCs may have potential clinical use in regenerative medicine

    • 2015-10-06
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    Healthspan-Extending Activity of Human Amniotic Membrane- and Adipose Tissue-Derived Stem Cells in F344 Rats. Stem Cells Translational Medicine

    요약 : 
    Ageing brings about the progressive decline in cognitive function and physical activity, along with losses of stem cell population and function. Although transplantation of muscle derived stem/progenitor cells extended healthspan and lifespan of progeria mice, such effects in normal animals were not confirmed. Human amniotic membrane-derived mesenchymal stem cells (AMMSCs) or adipose tissue-derived mesenchymal stem cells (ADMSCs) (1 × 106 55 cells/rat) were intravenously transplanted to 10-month-old male F344 rats once a month for their life-long periods. Transplantation of AMMSCs and ADMSCs improved cognitive and physical functions of naturally-ageing rats, extending lifespan by 23.4% and 31.3%, respectively. The stem cell therapy increased the concentration of acetylcholine, and recovered neurotrophic factors in the brain and muscles, leading to restoration of microtubule-associated protein 2, cholinergic and dopaminergic nervous systems, microvessels, muscle mass as well as antioxidative capacity. The results indicate that repeated transplantation of AMMSCs and ADMSCs elongate both healthspan and lifespan, which could be a starting point for anti-ageing or rejuvenation effects of allogeneic or autologous stem cells with minimum immune rejection.

    • 2015-10-06
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    Epithelial to mesenchymal transition enhances the cardioprotective capacity of human amniotic epithelial cells

    요약 : 
    The amniotic epithelium consists of cells exhibiting mature epithelial cell characteristics, but also varying degrees of stemness. We tested the hypothesis that induction of epithelial-to-mesenchymal transition (EMT) in
    amniotic epithelial cells (AECs) derived from human placenta enhances their capacity to support the ischemic myocardium. In response to incubation with transforming growth factor-b1 (TGF-b1) protein, AECs lost their
    cobblestone morphology and acquired a fibroblastoid shape, associated with downregulation of E-cadherin, upregulation of N-cadherin, Akt phosphorylation, and intracellular periostin translocation. EMT–AECs displayed greatly enhanced mobility and secreted gelatinase activity compared with naive AECs. The surface presentation of CD105 and CD73 decreased, and RNA microarray analysis mirrored the loss of epithelial characteristics and transcriptional profile. Unmodified AECs and EMT–AECs were then injected intramyocardially in fully immunocompetent mice after permanent LAD ligation, and heart function was followed by MRI as well as 2D speckle tracking echocardiography after 4 weeks. EMT–AEC-treated infarct hearts displayed better global systolic function and improved longitudinal strain rate in the area of interest. Although no signals of human cells were detectable by histology, infarct size was smaller in EMT–AEC-treated hearts, associated with fewer TUNEL-positive cells and upregulation of periostin, while blood vessel density was increased in both ACE- and EMT–AEC-treated hearts. We conclude that EMT enhances the cardioprotective effects of human AECs

    • 2015-10-06
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    Anti-Wrinkle Effects of Adipose Tissue-Derived Mesenchymal Stem Cells in a UV-Irradiated Hairless Mouse Model

    요약 : 
    The potential anti-wrinkle effects of adipose tissue-derived mesenchymal stem cells (AdMSCs) have recently been reported. In the present study, we demonstrated a protective effect of AdMSCs in a UV irradiationinduced
    hairless mouse wrinkle model. Mice treated with AdMSCs showed improvements in skin erythema index, moisture capacity and transepidermal water loss. The AdMSC-treated mice showed reduced wrinkle area, as measured by gross observation and replica image analysis, and had a thinner epidermis and increased collagen and elastic fiber content, compared with the control group, as measured by histopathological examination. These results suggested that AdMSCs may be effective in the treatment of wrinkles. The activity of AdMSCs was further confirmed by the reduction in MMP-3 mRNA expression in the skin of the AdMSC treated animals, which suggests that AdMSCs can protect the collagen from degradation induced by UV irradiation. Further studies will investigate detailed mechanisms of action of AdMSCs, with particular focus on AdMSC-derived secretory factors

    • 2015-10-06
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    Anti-wrinkle Effect of AdMSCs-CM in Photoaging Skin Model of Hairless Mice

    요약 : 
    The purpose of this study was to investigate the anti-wrinkle effect of adipose-derived mesenchymal stem cell-conditioned medium (AdMSCs-
    CM) in a UVB irradiation-induced hairless mouse wrinkle model. Mice treated with AdMSCs-CM showed improvements in transepidermal water loss (TEWL), water capacity and erythema index of skin. The AdMSCs-CM treated mice showed reduced total wrinkle area, number, length and depth, as measured by gross observation and replica image analysis, and had a thinner epidermis and increased collagen and elastic fiber content, compared with the UV control group, as measured by histopathological examination. The activity of AdMSCs-CM was further confirmed by the reduction in MMP-3 mRNA expression and MMP-2, 9 protein activity in the skin of the AdMSCs CM treated animals. These results suggested that AdMSCs-CM may be effective in the treatment of wrinkles by protecting of the collagen from degradation induced by UV irradiation. Further studies about detailed mechanisms of action of AdMSCs-CM will be needed

    • 2015-10-06
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    Amniotic membrane extract-loaded double-layered wound dressing: evaluation of gel properties and wound healing

    요약 : 
    The conservative single-layered wound dressing system is decomposed when mixed in polyvinyl alcohol (PVA) solution, which means it cannot be used with a temperature-sensitive drug. The goal of this investigation was to make an amniotic membrane extract (AME)-loaded double-layered wound dressing with an improved healing result compared to the conservative single-layered wound dressing systems. The double-layered wound dressing was developed with PVA/sodium alginate using a freeze–melting technique; one layer was PVA layer and the other was the drug-loaded sodium alginate layer. Its gel properties were assessed compared to single-layered wound dressings. Moreover, in vivo wound-healing effects and histopathology were calculated compared to commercial products. The double-layered wound dressing gave a similar gel fraction and Young’s module as single-layered wound bandages developed with only PVA, and a similar inflammation ability and WVTR as single-layered wound dressings developed with PVA and sodium alginate. Our data indicate that these double-layered wound bandages were just as swellable, but more elastic and stronger than single-layered wound dressings comprised of the same polymers and quantities, possibly giving an acceptable level of moisture and accumulation of exudates in the wound zone. Compared to the commercial product, the double-layered wound dressing comprising 6.7% PVA, 0.5% sodium alginate and 0.01% AME significantly enhanced the wound-healing effect in the wound-healing test. Histological investigations showed that superior full-thickness wound-healing effects compared to the commercial product. Therefore, the double-layered wound dressing would be an outstanding wound-dressing system with improved wound healing and good gel property

    • 2015-10-06
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    Therapeutic lymphangiogenesis using stem cell and VEGF-C hydrogel

    요약 : 
    Lymphedema is a manifestation of lymphatic system insufficiency. It arises from primary lymphatic dysplasia or secondary obliteration after lymph node dissection or irradiation. Although improvement of swelling can be achieved by comprehensive non-operative therapy, treatment of this condition requires lifelong care and good compliance. Recently molecular-based treatments using VEGF-C have been investigated by several researchers. We designed the present study to determine whether the therapeutic efficacy of implanted human adipose-derived stem cells (hADSCs) could be improved by applying a gelatin hydrogel containing VEGF-C (VEGF-C hydrogel) to the site of tissue injury in a lymphedema mouse model. Four weeks after the operation, we evaluated edema and determined lymphatic vessel density at various post-operative time points. Mice treated with hADSCs and VEGF-C hydrogel showed a significantly decreased dermal edema depth compared to the groups of mice that received hADSCs only or VEGF-C hydrogel only. Immunohistochemical analysis also revealed that the hADSC/VEGF-C hydrogel group showed significantly greater lymphatic vessel regeneration than all the other groups. hADSCs were detected in the implantation sites of all mice in the hADSC/VEGF-C group, and exhibited a lymphatic endothelial differentiation phenotype as determined by co-staining PKH-labeled hADSCs for the lymphatic marker LYVE-1. Our results suggest that co-administration of hADSCs and VEGF-C hydrogel has a substantial positive effect on lymphangiogenesis

    • 2015-10-06
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    Systemic transplantation of human adipose tissue-derived mesenchymal stem cells for the regeneration of irradiation-induced salivary gland damage

    요약 : 
    Objectives: Cell-based therapy has been reported to repair or restore damaged salivary gland (SG) tissue after irradiation. This study was aimed at determining whether systemic administration of human adipose-derived mesenchymal stem cells (hAdMSCs) can ameliorate radiation-induced SG damage.
    Methods: hAdMSCs (16106) were administered through a tail vein of C3H mice immediately after local irradiation, and then this infusion was repeated once a week for 3 consecutive weeks. At 12 weeks after irradiation, functional evaluations were conducted by measuring salivary flow rates (SFRs) and salivation lag times, and histopathologic and immunofluorescence histochemistry studies were performed to assay microstructural changes, apoptosis, and proliferation indices. The engraftment and in vivo differentiation of infused hAdMSCs were also investigated, and the transdifferentiation of hAdMSCs into amylase-producing SG epithelial cells (SGCs) was observed in vitro using a co-culture system.

    • 2015-10-06
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    Anti-tumor effects of canine adipose tissue-derived mesenchymal stromal cell-based interferon-β gene therapy and cisplatin in a mouse melanoma model.

    요약 :
    Background aims. Adipose tissue (AT)-derived mesenchymal stromal cells (MSC) (AT-MSC) represent a novel tool for delivering therapeutic genes to tumor cells. Interferon (IFN)- β is a cytokine with pleiotropic cellular functions, including anti-proliferative, immunomodulatory and anti-angiogenic activities. The purpose of this study was to engineer canine AT-MSC (cAT-MSC) producing IFN- β and to evaluate the anti-tumor effect of cAT-MSC – IFN- β combined with cisplatin in mouse melanoma model. Methods. cAT-MSC engineered to express mouse IFN- β were generated using a lentiviral vector (cAT-MSC – IFN- β ) and the secreted IFN- β -induced inhibition of tumor cell growth and apoptosis on B16F10 cells was investigated in vitro prior to in vivo studies. Melanoma-bearing mouse was developed by injecting B16F10 cells subcutaneously into 6-week-old C57BL/6 mice. After 14 days, cisplatin (10 mg/kg) was injected intratumorally, and 3 days later the engineered cAT-MSC were injected subcutaneously every 3 days to death. Tumor volume and survival times were measured.
    Results. The combination treatment of cAT-MSC – IFN- β with cisplatin was more effective in inhibiting the growth of melanoma and resulted in signifi cantly extended survival time than both an unengineered cAT-MSC – cisplatin combination group and a cisplatin-alone group. Interestingly, subcutaneously injected cAT-MSC – IFN- β were migrated to tumor sites. Conclusions. Our data suggest that canine AT-MSC could serve as a powerful cell-based delivery vehicle for releasing therapeutic proteins to tumor lesions. Maximal anti-tumor effects were seen when this therapy was combined with a DNAdamaging chemotherapeutic agent. This study demonstrates the possible applicability of AT-MSC-mediated IFN- β in treating canine and human cancer patients

    • 2015-10-06
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    Anti-tumor effect of adipose tissue derived-mesenchymal stem cells expressing interferon-β and treatment with cisplatin in a xenograft mouse model for canine melanoma

    요약 : 
    Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are attractive cell-therapy vehicles for the delivery of anti-tumor molecules into the tumor microenvironment. The innate tropism of AT-MSCs for tumors has important implications for effective cellular delivery of anti-tumor molecules, including cytokines, interferon, and pro-drugs. The present study was designed to determine the possibility that the combination of stem cell-based gene therapy with low-dose cisplatin would improve therapeutic efficacy against canine melanoma. The IFN-b transduced canine AT-MSCs (cAT-MSC-IFN-b) inhibited the growth of LMeC canine melanoma cells in direct and indirect in vitro co-culture systems. In animal experiments using BALB/c nude mouse xenografts, which developed by injecting LMeC cells, the combination treatment of cAT-MSC-IFN-b and low-dose cisplatin significantly reduced tumor volume compared with the other treatment groups. Fluorescent microscopic analysis with a TUNEL (terminal deoxynucleotidyl transferase-mediated nick-end labeling) assay of tumor section provided evidence for homing of cAT-MSC-IFN-b to the tumor site and revealed that the combination treatment of cAT-MSC-IFN-b with low-dose cisplatin induced high levels of cell apoptosis. These findings may prove useful in further explorations of the application of these combined approaches to the treatment of malignant melanoma and other tumors

    • 2015-10-06
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    Therapeutic effect of adipose-derived stem cells and BDNF-immobilized PLGA membrane in a rat model of cavernous nerve injury

    요약 : 
    Introduction: Cavernous nerve injury is the main reason for post-prostatectomy erectile dysfunction (ED). Stem cell and neuroprotection therapy are promising therapeutic strategy for ED. Aim. To evaluate the therapeutic efficacy of adipose-derived stem cells (ADSCs) and brain-derived neurotrophic factor (BDNF) immobilized Poly-Lactic-Co-Glycolic (PLGA) membrane on the cavernous nerve in a rat model of post-prostatectomy ED. Methods. Rats were randomly divided into five groups: normal group, bilateral cavernous nerve crush injury (BCNI) group, ADSC (BCNI group with ADSCs on cavernous nerve) group, BDNF-membrane (BCNI group with BDNF/PLGA membrane on cavernous nerve) group, and ADSC/BDNF-membrane (BCNI group with ADSCs covered with BDNF/PLGA membrane on cavernous nerve) group. BDNF was controlled-released for a period of 4 weeks in a BDNF/PLGA porous membrane system. Main Outcome Measures. Four weeks after the operation, erectile function was assessed by detecting the ratio of intra-cavernous pressure (ICP)/mean arterial pressure (MAP). Smooth muscle and collagen content were etermined by Masson’s trichrome staining. Neuronal nitric oxide synthase (nNOS) expression in the dorsal penile nerve was detected by immunostaining. Phospho-endothelial nitric oxide synthase (eNOS) protein expression and cyclic guanosine monophosphate (cGMP) level of the corpus cavernosum were quantified byWestern blotting and cGMP assay, respectively.

    • 2015-10-06
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    Effect of an adipose-derived stem cell and nerve growth factor-incorporated hydrogel on recovery of erectile function in a rat model of cavernous nerve injury.

    요약 : 
    Postprostatectomy erectile dysfunction (ED) is the major problem for patients with clinically localized prostate cancer. Recently, gene and stem cell-based therapy of the corpus cavernosum has been attempted for postprostatectomy ED, but those therapies are limited by rapid blood flow and disruption of the normal architecture of the corpus cavernosum. In this study, we attempted to regenerate the damaged cavernous nerve (CN), which is the main cause of ED. We investigated the effectiveness of human adipose-derived stem cell (hADSC) and nerve growth factor-incorporated hyaluronic acid-based hydrogel (NGF-hydrogel) application on the CN in a rat model of bilateral cavernous nerve crush injury. Four weeks after the operation, erectile function was assessed by detecting the intracavernous pressure (ICP)/arterial pressure level by CN electrostimulation. The ICP was significantly increased by application of hADSC with NGF-hydrogel compared to the other experimental groups. CN and penile tissue were collected for histological examination. PKH-26 labeled hADSC colocalized
    with beta III tubulin were shown in CN tissue sections. hADSC/NGF-hydrogel treatment prevented smooth muscle atrophy in the corpus cavernosum. In addition, the hADSC/NGF-hydrogel group showed increased endothelial nitric oxide synthase protein expression. This study suggests that application of hADSCs with NGFhydrogel on the CN might be a promising treatment for postprostatectomy ED

    • 2015-10-06
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    Journey of mesenchymal stem cells for homing: strategies to enhance efficacy and safety of stem cell therapy

    요약 : 
    Humanmesenchymal stem cells (MSCs) communicate with other cells in the human body and appear to “home” to areas of injury in response to signals of cellular damage, known as homing signals. This review of the state of current research on homing of MSCs suggests that favorable cellular conditions and the in vivo environment facilitate and are required for the migration of MSCs to the site of insult or injury in vivo. We review the current understanding of MSC migration and discuss strategies for enhancing both the environmental and cellular conditions that give rise to effective homing of MSCs. This may allow MSCs to quickly find and migrate to injured tissues, where they may best exert clinical benefits resulting from improved homing and the presence of increased numbers of MSCs.

    • 2015-10-06
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    Safety of intravenous infusion of human adipose tissue-derived mesenchymal stem cells in animals and humans.

    요약 :
    Adipose tissue-derived mesenchymal stem cells (AdMSCs) represent an attractive and ethical cell source for stem cell therapy. With the recent demonstration of MSC homing properties, intravenous applications of MSCs
    to cell-damaged diseases have increased. In the present study, the toxicity and tumorigenicity of human AdMSCs (hAdMSCs) were investigated for clinical application. Culture-expanded hAdMSCs showed the typical
    appearance, immunophenotype, and differentiation capacity of MSCs, and were genetically stable at least 12 passages in culture. Cells suspended in physiological saline maintained their MSC properties in a cold storage
    condition for at least 3 days. To test the toxicity of hAdMSCs, different doses of hAdMSCs were injected intravenously into immunodeficient mice, and the mice were observed for 13 weeks. Even at the highest cell
    dose (2.5 · 108 cells/kg body weight), the SCID mice were viable and had no side effects. A tumorigenicity test was performed in Balb/c-nu nude mice for 26 weeks. Even at the highest cell dose (2 · 108 MSCs/kg), no
    evidence of tumor development was found. In a human clinical trial, 8 male patients who had suffered a spinal cord injury > 12 months previous were intravenously administered autologous hAdMSCs (4 · 108 cells) one
    time. None of the patients developed any serious adverse events related to hAdMSC transplantation during the 3-month follow-up. In conclusion, the systemic transplantation of hAdMSCs appears to be safe and does not
    induce tumor development

    • 2015-10-06
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    Recloned dogs derived from adipose stem cells of a transgenic cloned beagle

    요약 : 
    A number of studies have postulated that efficiency in mammalian cloning is inversely correlated with donor cell differentiation status and may be increased by using undifferentiated cells as nuclear donors. Here, we attempted the recloning of dogs by nuclear transfer of canine adipose tissue-derived mesenchymal stem cells (cAd-MSCs) from a transgenic cloned beagle to determine if cAd-MSCs can be a suitable donor cell type. In order to isolate cAd-MSCs, adipose tissues were collected from a transgenic cloned beagle produced by somatic cell nuclear transfer (SCNT) of canine fetal fibroblasts modified genetically with a red fluorescent protein (RFP) gene. The cAd-MSCs expressed the RFP gene and cell-surface marker characteristics of MSCs including CD29, CD44 and thy1.1. Furthermore, cAd-MSCs underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. In order to investigate the developmental potential of cAd-MSCs, we carried out SCNT. Fused-couplets (82/109, 75.2%) were chemically activated and transferred into the uterine tube of five naturally estrus-synchronized surrogates. One of them (20%) maintained pregnancy and subsequently gave birth to two healthy cloned pups. The present study demonstrated for the first time the successful production of cloned beagles by nuclear transfer of cAd-MSCs. Another important outcome of the present study is the successful recloning of RFP-expressing transgenic cloned beagle pups by nuclear transfer of cells derived from a transgenic cloned beagle. In conclusion, the present study demonstrates that adipose stem cells can be a good nuclear donor source for dog cloning

    • 2015-10-06
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    Generation of transgenic dogs that conditionally express green fluorescent protein

    요약 : 
    We report the creation of a transgenic dog that conditionally expresses eGFP (enhanced green fluorescent protein) under the regulation of doxycycline. Briefly, fetal fibroblasts infected with a Tet-on eGFP
    vector were used for somatic cell nuclear transfer. Subsequently reconstructed oocytes were transferred to recipients. Three clones having transgenes were born and one dog was alive. The dog showed all features of inducible expression of eGFP upon doxycycline administration, and successful breeding resulted in eGFPpositive puppies, confirming stable insertion of the transgene into the genome. This inducible dog model
    will be useful for a variety of medical research studies

    • 2015-10-06
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    Generation of red fluorescent protein transgenic dogs.

    요약 :
    Dogs (Canis familiaris) share many common genetic diseases with humans and development of disease models using a transgenic approach has long been awaited. However, due to the technical difficulty in
    obtaining fertilizable eggs and the unavailability of embryonic stem cells, no transgenic dog has been generated. Canine fetal fibroblasts were stably transfected with a red fluorescent protein (RFP) gene-expressing
    construct using retrovirus gene delivery method. Somatic cell nuclear transfer was then employed to replace the nucleus of an oocyte with the nucleus of the RFPfibroblasts. Using this approach, we produced the first
    generation of transgenic dogs with four female and two male expressing RFP

    • 2015-10-06
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    Quantification of early adipose-derived stem cell survival: comparison between sodium iodide symporter and enhanced green fluorescence protein imaging.

    요약 : 
    Objective: Strategies to overcome the problem of extensive early stem cell loss following transplantation requires a method to quantitatively assess their efficacy. This study compared the ability of sodium/iodide symporter (NIS) and enhanced green fluorescent protein (EGFP) imaging to monitor the effectiveness of treatments to enhance early stem cell survival.
    Methods: Human adipose-derived stem cells (ADSCs) transduced with an adenoviral vector to express both NIS and EGFP were mixed with culture media (control), matrigel (matrigel group) or pro-survival cocktail
    (PSC group), and 5×106 cells were injected into thigh muscles of C57BL/6 mice. Animals underwent serial optical imaging and 99mTcO4 – scintigraphy. Image-based EGFP fluorescence and 99mTcO4 – uptake was measured by region-of-interest analysis, and extracted tissues were measured for 99mTc activity. Fluorescent intensity measured from homogenized muscle tissue was used as reference for actual amount of viable ADSCs.
    Results: ADSCs were efficiently transduced to express EGFP and NIS without affecting proliferative capacity. The absence of significant apoptosis was confirmed by annexin V FACS analysis and Western blots for activated caspase-3. Both fluorescence optical imaging and 99mTcO4 – scintigraphy visualized implanted cells in living mice for up to 5days. However, optical imaging displayed large variations in fluorescence intensity, and thus failed to detect difference in cell survival between groups or its change over time. In comparison, 99mTcO4 -scintigraphy provided more reliable assessment of within-in group donor cell content as well as its temporal change. As a result, NIS imaging was able to discern beneficial effects of matrigel and pro-survival cocktail treatment on early ADSC survival, and provided quantitative measurements that correlated to actual donor cell content within implanted tissue.
    Conclusion: NIS reporter imaging may be useful for noninvasively assessing the efficacies of strategies designed to improve early survival of transplanted stem cell

    • 2015-10-06
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    Improved viability and activity of neutrophils differentiated from HL-60 cells by co-culture with adipose tissue-derived mesenchymal stem cells

    요약 :
    Neutropenia is a principal complication of cancer treatment. We investigated the supportive effect of adipose tissue-derived mesenchymal stem cells (AD-MSCs) on the viability and function of neutrophils. Neutrophils were derived from HL-60 cells by dimethylformamide stimulation and cultured with or without AD-MSCs under serum-starved conditions to evaluate neutrophil survival, proliferation, and function.
    Serum starvation resulted in the apoptosis of neutrophils and decreased cell survival. The co-culture of neutrophils and AD-MSCs resulted in cell survival and inhibited neutrophil apoptosis under serumstarved conditions. The survival rate of neutrophils was prolonged up to 72 h, and the expression levels of interferon (IFN)-a, granulocyte colony-stimulating factor (G-CSF), granulocyte–macrophage colonystimulating factor, and transforming growth factor (TGF)-b in AD-MSCs were increased after co-culture with neutrophils. AD-MSCs promoted the viability of neutrophils by inhibiting apoptosis as well as enhancing respiratory burst, which could potentially be mediated by the increased expression of IFNa, G-CSF, and TGF-b. Thus, we conclude that the use of AD-MSCs may be a promising cell-based therapy for increasing immunity by accelerating neutrophil function

    • 2015-10-06
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    Comparison of human muscle-derived stem cells and human adipose-derived stem cells in neurogenic trans-differentiation.

    요약 : 
    Purpose: Erectile dysfunction (ED) remains a major complication from cavernous nerve injury during radical prostatectomy. Recently, stem cell treatment for ED has been widely reported. This study was conducted to investigate the availability, differentiation into functional cells, and potential of human muscle-derived stem cells (hMDSCs) and human adipose-derived stem cells (hADSCs) for ED treatment. Materials and Methods: We compared the neural differentiation of hMDSCs and hADSCs. Human muscle and adipose tissues were digested with collagenase, followed
    by filtering and centrifugation. For neural induction, isolated hMDSCs and hADSCs were incubated in neurobasal media containing forskolin, laminin, basic-fibroblast growth factor, and epidermal growth factor for 5 days. Following neural induction, hMDSCs and hADSCs were differentiated into neural cells, including neurons and glia, in vitro.
    Results: In neural differentiated hMDSCs (d-hMDSCs) and differentiated hADSCs (d-hADSCs), neural stem cell marker (nestin) showed a significant decrease by immunocytochemistry, and neuronal marker (β-tubulin III) and glial marker (GFAP) showed a significant increase, compared with primary hMDSCs and hADSCs. Real-time chain reaction analysis and Western blotting demonstrated significantly elevated levels of mRNA and protein of β-tubulin III and GFAP in d-hADSCs compared with d-hMDSCs.
    Conclusions: We demonstrated that hMDSCs and hADSCs can be induced to undergo phenotypic and molecular changes consistent with neurons. The neural differentiation capacity of hADSCs was better than that of hMDSCs

    • 2015-10-06
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    In vitro migration capacity of human adipose tissue-derived mesenchymal stem cells reflects their expression of receptors for chemokines and growth factors.

    요약 : 
    The homing properties of adipose tissue-derived mesenchymal stem cells (AdMSCs) have stimulated intravenous applications for their use in stem cell therapy. However, the soluble factors and corresponding cellular
    receptors responsible for inducing chemotaxis of AdMSCs have not yet been reported. In the present study, the migration capacity of human AdMSCs (hAdMSCs) toward various cytokines or growth factors (GFs) and the expression of their receptors were determined. In a conventional migration assay, PDGF-AB, TGF-β1, and TNF-α showed the most effective chemoattractant activity. When AdMSCs were preincubated with various chemokines or GF, and then allowed to migrate toward medium containing 10% FBS, those preincubated with TNF-α showed the highest migratory activity. Next, hAdMSCs were either preincubated or not with TNF-α, and allowed to migrate in response to various GFs or chemokines. Prestimulation with TNF-α increased the migration activity of hAdMSCs compared to unstimulated hAdMSCs. When analyzed by FACS and RT-PCR methods, hAdMSCs were found to express C-C chemokine receptor type 1(CCR1), CCR7, C-X-C chemokine receptor type 4 (CXCR4), CXCR5, CXCR6, EGF receptor, fibroblast growth factor receptor 1, TGF-β receptor 2, TNF receptor superfamily member 1A, PDGF receptor A and PDGF receptor B at both the protein and the mRNA levels. These results indicate that the migration capacity of hAdMSCs is controlled by various GFs and chemokines. Prior in vitro modulation of the homing capacity of hAdMSCs could stimulate their movement into injured sites in vivo when administered intravenously, thereby improving their therapeutic potential.

    • 2015-10-06
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    Effects of expanded human adipose tissue-derived mesenchymal stem cells on the viability of cryopreserved fat grafts in the nude mouse.

    요약 :
    Adipose-derived mesenchymal stem cells (AdMSCs) augment the ability to contribute to microvascular remodeling in vivo and to modulate vascular stability in fresh fat grafts. Although cryopreserved adipose tissue is frequently used for soft tissue augmentation, the viability of the fat graft is poor. The effects of culture-expanded human adipose tissue-derived mesen-chymal stem cells (hAdMSCs) on the survival and quality of the cryopreserved fat graft were determined. hAdMSCs from the same donor were mixed with fat tissues cryopreserved at –70°C for 8 weeks and injected subcutaneously into 6-week-old BALB/c-nu nude mice. Graft volume and weight were measured, and histology was evaluated 4 and 15 weeks post-transplantation. The hAdMSC-treated group showed significantly enhanced graft volume and weight. The histological evaluation demonstrated significantly better fat cell integrity compared with the vehicle-treated control 4 weeks post-transplantation. No significant dif-ference in graft weight, volume, or histological parameters was found among the groups 15 weeks post-transplantation. The hAdMSCs enhanced the survival and quality of transplanted cryopreserved fat tissues. Cultured and expanded hAdMSCs have reconstructive capacity in cryopreserved fat grafting by increasing the number of stem cells

    • 2015-10-06
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    Comparison of cytokine expression in mesenchymal stem cells from human placenta, cord blood, and bone marrow.

    요약 : 
    Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation into lineages of mesenchymal tissues that are currently under investigation for a variety of therapeutic applications. The purpose of this study was to compare cytokine gene expression in MSCs from human placenta, cord blood (CB) and bone marrow (BM). The cytokine expression profiles of MSCs from BM, CB and placenta (amnion, decidua) were compared by proteome profiler array analysis. The cytokines that were expressed differently, in each type of MSC, were analyzed by real-time PCR. We evaluated 36 cytokines. Most types of MSCs had a common expression pattern including MIF (GIF, DER6), IL-8 (CXCL8), Serpin E1 (PAI-1), GROα(CXCL1), and IL-6. MCP-1, however, was expressed in both the MSCs from the BM and the amnion. sICAM-1 was expressed in both the amnion and decidua MSCs. SDF-1 was expressed only in the BM MSCs. Real-time PCR demonstrated the expression of the cytokines in each of the MSCs. The MSCs from bone marrow, placenta (amnion and decidua) and cord blood expressed the cytokines differently. These results suggest that cytokine induction and signal transduction are different in MSCs from different
    tissues

    • 2015-10-06
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    New Isolation Technique and Culture System for Clinical Applications of Human Amniotic Epithelial Stem Cells.

    요약 : 
    태반은 성체줄기세포의 보고이다. 특히 양막상피세포는 배아줄기세포의 줄기세포 능력을 나타내는 세포 표면표시자들을 그대로 발현하는 줄기세포로 알려져 있다. 하지만 상피세포를 실험실에서 지지세포 없이 대량 증식 배양하는 것은 상피세포가 가지고 있는 내인성 성격으로 인해 어렵다. 본 연구에서는 디티오트레이톨(Dithiothreitol; DTT)과 ROCK 저해제(Rho-associated kinase inhibitor)를 이용하여 양막상피세포를 분리하고 배양하는데 있어서 임상적용이 가능한 수준의 세포를 얻었고, 최적의 세포상태를 유지하였다. 본 연구에서 분리배양된 양막상피세포는 상피세포의 특성과 줄기세포의 특성을 발현하였다. 결론적으로 줄기세포 치료를 이용한 재생의학의 관점에서 인간태반 유래 양막상피줄기세포는 아무런 윤리적인 논란을 일으키지 않는 주요한 줄기세포 치료제의 재료로서 여러 가지 질병 치료에 사용될 수 있을 것이다.

    • 2015-10-06
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    Comparison of neural cell differentiation of human adipose mesenchymal stem cells derived from young and old ages

    요약 : 
    최근 골수와 혈액으로 유래된 중간엽 줄기세포와 비슷한 능력을 가지는 것으로 알려진 지방 유래 중간엽줄기세포가 새로운 세포 치료제로 떠오르고 있다. 하지만 줄기세포를 이용하여 치료하려는 질병은 나이가 들어감에 따라 발병하는 퇴행성 질환들이 대부분인데, 노화가 진행됨에 따라 줄기세포의 능력이 차이가 있다고 알려져 있다. 이에 본 연구에서는 노화가 일어남에 따라 발생되는 신경성 질환을 자가 유래 지방 중간엽 줄기세포를 이용하여 치료함에 있어서 노화가 진행됨에 따라 얻어진 지방줄기세포가 세포학적으로 변화는 없는지에 대해 줄기세포 성장능, 생존율과 신경세포로의 분화유도 능력을 비교하였다. 30대, 40대, 50대에서 사람 지방 유래 줄기세포를 분리 배양하여 연령별 계대에 따른 세포수와 생존율을 측정하고, 줄기세포 성장능력을 비교 분석하였고, 지방 줄기세포를 신경세포 배양 조건 하에서 10일 동안 배양하여 신경 분화능력을 연령별로 비교하였다. 실험결과, 세포수와 생존율, 세포 모양이 연령과 계대별에 의해 차이가 없다는 것을 확인하였다. 신경 분화 후 면역형광염색법을 통해 분석한 결과, 연령에 따른 신경 분화능력의 차이가 관찰되지 않았다. 분자 유전적학으로 신경세포 마커의 발현을 mRNA 수준에서 분석한 결과, 연령별 간의 차이가 몇 개의 유전자 발현을 제외하고는 차이가 발견되지 못했다. 하지만 계대가 진행될수록 50대군의 줄기세포에서 MAP2와 Sox2의 mRNA 발현이 30대군의 줄기세포에 비해 상대적으로 낮게 발현됨이 확인되었다. 결론적으로 자가 지방 중간엽 줄기세포의 신경세포 분화능력이 연령에 상관없이 차이가 없음이 관찰되었으며, 이는 나이 든 사람으로부터 얻어진 지방 줄기세포도 젊은 사람에서 얻어진 세포와 마찬가지 능력으로 자가 세포 치료제로 사용될 수 있다는 점을 말해주고 있다.

    • 2015-10-06
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    HOXC10 as a potential marker for discriminating between amnion- and decidua-derived mesenchymal stem cells.

    요약 :
    The HOX family of genes plays a fundamental role in the morphogenesis of vertebrate embryonic cells. HOX genes are thought to be important for the regulation of stem cells. We investigated HOX gene expression in
    mesenchymal stem cells (MSCs) from human placentas. We isolated MSCs from human placentas and confirmed stemness by fluorescence-activated cell sorting (FACS) analysis and differentiation studies. Using reverse
    transcription PCR, mRNA expression of 39 Class I HOX genes was measured in the MSCs. The expression of HOXB6, C4, C8, C10, D3, D4, and D10 were measured by Western blot analysis. HOXC10 was expressed in 10
    of 10 amnion-derived MSCs but in only 2 of 10 decidua-derived MSCs. HOXC4 and D10 were expressed in 100% of both amnion-derived MSCs and deciduas-derived MSCs. HOXD4 was silent in all amnion-derived MSCs and deciduas-derived MSCs (n¼10). HOX gene activation patterns might be a useful indicator for the detection of MSCs of different tissue origins. We demonstrated that HOXC10 is a gene that may discriminate between
    amnion-derived MSCs and decidua-derived MSCs

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