【摘要】 目的:探讨人脂肪基质干细胞体外诱导为血管平滑肌细胞的可能性。方法:应用酶消化法从人脂肪组织获得单个核细胞,基础培养液培养,经传代后,第一代细胞用于诱导分化实验。未诱导组培养液为基础培养液,诱导组培养液在基础培养液内添加诱导因子PDGF-BB(50ng/mL)、TGF-β1(5ng/mL),诱导14d后,光镜下观察诱导细胞形态学的改变;免疫荧光和RT-PCR方法检测平滑肌细胞特异标记的表达情况;流式细胞仪检测诱导阳性率。结果:脂肪基质干细胞在特定细胞因子诱导作用下,细胞生长呈现与平滑肌细胞类似的"峰-谷"生长模式,并表达α-SM-Actin、SM-MHC、Calponin、SM-22α等平滑肌特异性标记;诱导组阳性率与未诱导组差异有统计学意义(P<0.01)。结论:PDGF-BB、TGF-β1可诱导人脂肪基质干细胞向血管平滑肌细胞表型分化。更多还原

【Abstract】 Objective To study the feasibility of human adipose derived stromal cells (ADSCs) induced into smooth muscle cells in vitro. Methods The mononuclear cells in human adipose were separated by collagenase treatment and seeded on culture dishes.Cells were cultured in M-199 plus 10% FBS.When reaching confluence,the cells were subcultured and PDGF-BB and TGF-β1 were added at the passage 1 to enhance the smooth muscle cells’ phenotype.Cells were cultured under the inducing medium for 14 days.Cell morphology was observed under microscopy.Cellular immunofluorescence and RT-PCR were used to determine the expression of smooth muscle cell markers of the post-induced cells.FACS was used to examine the positive rate of induced team. Results After subculture,the induced cells exhibited "Hill and Valley" morphology,and expressed SM cell-specific markers including α-SM-Actin,SM-MHC,calponin and SM-22α.The positive rates were remarkable higher than those in uninduced cells. Conclusion The human ADSCs can be induced to express vascular smooth muscle markers.更多还原