人类脂肪来源成体干细胞体外定向成软骨的诱导实验

Objective To investigate the feasibility of inducing human adipose derived adult stem cells (hADASc) into chondrocytes by gene transfection.Methods hADASc were cultured in vitro, and transfected with gene PGL3-TGF-pl, whose Luceferase activity (RUL value) was measured.PT-PCR was used to monitor the expression of tranfected hADASc.Anti collagen II immunohistochemical staining was performed in the experimental group (transfected hADASc) , active control group (untransfected hADASc cultured in chondrocytes induction medium) and negative control group (untransfected hADASc cultured in conventional medium).The immuno-staining image was analyzed with an automated imaging analysis system and computed for PU values.Results Nucleated tissue cells with features of stem cells were isolated from in vitro culture of human subcutaneous adipose tissue.The RUL value of the experimental group (9 212.583±315.240) was higher than that of the control group(317.000?0.710, P＜0.01).RT-PCR revealed that the transfected hADASc could express the TGF-β1.Anti collagen Ⅱ immunohistochemical staining was positive in experimental group and active control group, with PU values (13.864±2.416 and 13.637±2.548,respectively) statistically different from that of the negative control group (6.013 ±0.827, P＜0.05).Conclusions Nucleated tissue cells with features of stem cells can be isolated from culture of human subcutaneous adipose tissue in vitro.The PGL3-TGF-β1-transfected hADASc may express TGF-β1.The endogenous TGF-β1-induced hADASc produces collagen Ⅱ , with similar actions of exogenous TGF-β1.

Experimental induction of human adipose derived adult stem cells (hADASc) into chondrocytes in vitro