TGF-β1诱导绵羊肌源性干细胞向平滑肌分化的研究

发布时间：2018-03-27 18:05

本文选题：肌源性干细胞　切入点：转化生长因子β　出处：《中国妇产科临床杂志》2015年05期

【摘要】：目的通过改良差速贴壁法分离纯化绵羊MDSC,探索转化生长因子(TGF-β1)诱导绵羊肌源性干细胞分化为平滑肌的可能性。方法取成年绵羊股四头肌细胞采用改良差速贴壁法(modified preplate technique)进行优化、分离、纯化获得绵羊级源性干细胞(MDSC)。利用流式细胞仪鉴定CD44、CD34、CD31、CD45、CD14的表达;western bloting法检测干细胞标志物desmin表达情况鉴定MDSC。然后利用10 ng/ml TGF-β1诱导MDSC向平滑肌方向分化;随后利用Western bloting法检测诱导培养基处理的MDSC中平滑肌蛋白标志物a-SMA和calponin的表达;结果通过对改良差速贴壁法成功分离出成年绵羊的肌源性干细胞(MDSC)。流式细胞仪鉴定其干细胞标志物CD44、CD34阳性表达,同时CD31、CD45、CD14表达阴性。Western blotting检测MDSC中Desmin呈阳性表达。利用10 ng/ml TGF-β1成功诱导MDSC向平滑肌方向分化。诱导过程中,平滑肌特异蛋白a-SMA和calponin在诱导细胞中的表达逐步上升。结论本研究通过对差速贴壁法成功分离出成年雌性绵羊的肌源性干细胞(MDSC)。并首次利用TGF-β1诱导羊肌源性干细胞(MDSC)成功向平滑肌方向分化。
[Abstract]:Objective to study the possibility of inducing sheep myogenic stem cells to differentiate into smooth muscle cells by using modified differential adhesion method to isolate and purify sheep MDSC.Methods Adult sheep quadriceps femoris muscle cells were isolated and purified by modified differential adhesion method and induced by transforming growth factor TGF- 尾 1 (TGF- 尾 1). Modified preplate technique, Sheep grade derived stem cells were isolated and purified. The expression of CD44, CD41, CD41, CD45, CD14 was identified by flow cytometry. The expression of stem cell marker desmin was detected by western bloting. Then 10 ng/ml TGF- 尾 1 was used to induce the differentiation of MDSC into smooth muscle. The expression of smooth muscle protein markers a-SMA and calponin in MDSC treated with induction medium was detected by Western bloting assay. Results the myogenic stem cells of adult sheep were successfully isolated by modified differential adherence method. The positive expression of CD44-mCD34 was identified by flow cytometry. The expression of Desmin in MDSC was detected by Western blotting. 10 ng/ml TGF- 尾 1 was used to induce the differentiation of MDSC into smooth muscle. The expression of smooth muscle specific proteins a-SMA and calponin increased gradually. Conclusion in this study, the myogenic stem cells of adult female sheep were isolated successfully by differential adherence method. TGF- 尾 1 was used for the first time to induce the fine myogenic stem cells of sheep. MDSCs successfully differentiated into smooth muscle.
【作者单位】：广州医科大学附属第二医院妇科;北京协和医院妇科;
【分类号】：R711.2

【共引文献】