國家衛生研究院 NHRI:Item 3990099045/8383
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    題名: TGF-beta3 gene knockdown enhances stem cell properties in adipose derived stromal cells
    其他題名: TGF-β3 gene knockdown enhances stem cell properties in adipose derived stromal cells
    作者: Chen, TY;Lin, KMC
    貢獻者: Institute of Biomedical Engineering and Nanomedicine
    摘要: Problem:Notwithstanding many ongoing clinical trials of stem cell therapy targeting a variety of diseases are testing transfusion of stem cells originated from adult tissues such as adipose tissue, significant improvement after stem cell transplant has yet to be seen that likely because the need to raise the stem cell number as well as to improve the quality of transplantable cells is not met. Whereas both the number and quality of stem cells decline with aging, adipose derived stromal cells (ADSCs) have been considered as the most abundant mesenchymal stem cell-like cells of the elderly. A strategy to maximize the stemness of ADSCs will, therefore, enhance their therapeutic prospective and promote the realization of stem cell therapy. Background :TGF-β superfamily members such as transforming growth factor beta (TGF-βs), bone morphogenic proteins (BMPs) regulate a wide array of cellular processes including proliferation, differentiation, immune regulation. Through cross-talk between their respective pathways, TGF-βs and BMPs can modulate the activity of each other. Most of our knowledge was acquired from the studies on TGF-β1 and that other TGF-βs may display distinct, even opposing, functions has been shown elsewhere. Unlike TGF-β1, the role of TGF-β3 in many biological processes has not been extensively explored.Hypothesis: We proposed that knockdown of TGF-β3 expression may enhance the stem cell features and increase their immune-modulating capacity in ADSCs.Research:ADSCs isolated from wildtype and TGF-β3(+/-) mice carrying only one intact TGF-β3 allele were cultured. These ADSCs were subjected to adipogenic or osteogenic induction to compare their differentiation potential. To test the immune-modulating function, serum-free condition medium that was used to culture ADSCs for 24 h were collected, followed by concentrating step, and included in the stress challenge assays involving treating lung epithelial cells or fibroblasts with cigarette smoke extract (CSE). The cell death and the cytokines induced by CSE in these cells was used as the indication of cell injury, and the protection offered by ADSC condition medium was used to measure immune suppression capacity and pro-survival benefit of ADSCs. We also performed gene array analyses to study the involved pathways affected by TGF-β3 knockdown in ADSCs.Observations :Compared to wildtype, TGF-β3(+/-) ADSCs displayed significantly higher differentiation potential toward to both adipogenic and osteogenic lineage. The conditioned medium that was used to culture TGF-β3(+/-) ADSCs provided better protection against CSE-induced injury in lung epithelial cells and fibroblasts as indicated by an increased cell survival and by reduction of inflammatory cytokines in cells treated by TGF-β3(+/-) condition medium. Microarray data revealed interesting findings that losing one functional TGF-β3 allele in ADSCs not only reduced the level of TGF-β3 expression but also significantly decreased the expression of other TGF-β family members such as Bmp4. These results were confirmed further by experiments using ADSCs with TGF-β3 or BMP4 expression knockdown by siRNA transfections. Taken together, our study indicate that reduced TGF-β3 or BMP4 expression in ADSCs can simultaneously increase their differentiation potential toward multiple lineages and enhance their immune modulating capacity, both likely contributing to clinical success of stem cell therapy.
    日期: 2013-12
    關聯: World Stem Cell Summit Technology. 2013 Dec:Article number 27883.
    Link to: http://worldstemcellsummit.com/files/2013/app/PosterAbstracts2013.pdf
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