人脂肪间充质干细胞向表皮细胞表型转化的研究

目的:探索人脂肪间充质干细胞(adipose derived mesenchymal stem cells,ADSCs)向表皮细胞表型转化的方法。方法:以手术中剩余的人皮下脂肪组织为材料来源,利用胶原酶消化法分离ADSCs,流式细胞仪检测细胞表面标记CD29、CD90、CD105的表达,成脂诱导后油红O染色鉴定。实验组利用第四军医大学西京医院烧伤与皮肤外科实验室已成功构建的pc DNA3.1(+)/SP+EGF质粒经脂质体介导转染的Ha Cat细胞,与ADSCs在Transwell小室中共培养,对照组为ADSCs加入10%FBS培养基,14天后Realtime-PCR检测两组ADSCs中CK19、integrin-β的m RNA表达量。结果:实验组ADSCs中CK19、integrin-β的m RNA表达量较对照组明显升高,且差别有统计学意义。结论:转染EGF的Ha Cat细胞可诱导ADSCs向表皮细胞表型分化,从而为其成为组织工程理想的种子细胞提供了进一步的支持。

Cellular phenotype transdifferentiation of human adipose derived mesenchymal stem cells to epithelial cells

Objective To explore a method that inverts human adipose derived mesenchymal stem cells into epithelial cells. Methods The subcutaneous adipose tissue was collected from the operation. Primary ADSCs were isolated subsequently by the methods of stirring digestion within collagenase type I .The membrane antigens expression of cells,such as CD29,CD90, CD105 were detected through flowcytometry （ FCM ）. Adipocyte differentiation cells were stained for fat vacuoles using the Oil red O staining kit. The experiment group used pcDNA3.1 （＋）/SP-hEGF modified HaCat cells that was successfully created from our laboratory,and co-cultured with ADSCs in TranswelI.The control group was DMEM supplemented with fetal bovine serum for ADSCs.After inducing two weeks, the expression of CK19 and integrin-β mRNA were assessed by quantitative realtime PCR. Results The expression level of CK19 and integrin-β mRNA in the experiment group were obviously more than the control group, and there were significant statistical differences between two groups. Conclutions EGF transfected HaCat cells could induce ADSCs to invert to epithelial cells, which will further make ADSCs as ideal seed cells for tissue engineering.