分层共同培养诱导骨髓间充质干细胞向表皮细胞分化的研究

Study on the differentiation of mesenchymal stem cells into epidermal stem cells by the method of co-culture

目的探讨体外联合HaCaT细胞共同培养诱导骨髓间充质干细胞（MSCs）向表皮细胞分化的可行性。方法用聚碳酸酯细胞插入板分层后联合共同培养HaCaT细胞与MSCs，观察培养3、6、9d后的细胞形态，进行角蛋白（CK-19、CK-10）、整合素（仪6、B1）染色并用流式细胞仪统计细胞阳性率。结果共同培养后细胞形态变化明显，共培养3、6d后表皮细胞标志物CK-19、拍整合素、B1整合素免疫荧光染色阳性，细胞阳性率分别为9．3％、8．2％、11．5％和21．7％、34．1％、39．6％，CK10表达呈阴性；共培养9d后CK-19、α6整合素、B1整合素、表达较前减少，阳性率为12．2％、18．6％、16．3％，CK10出现阳性表达，细胞阳性率为10．7％。结论分层联合HaCaT细胞共同培养可以诱导骨髓干细胞向表皮细胞进行分化。

Objective To explore the feasibility of the differentiation of bone mesenchymal stem ceils （MSCs） into epidermal stem cells by the method of co-cuhure with HaCaT cell in vitro. Methods The morphologic changes of MSCs by the method of co-culture were observed with electronmicroscope after 3, 6, 9 d. lmmunohistochemical staining and flow cytometry were carried out to detect the positive staining of cytokeratin （CK-19, CK-10） and integrin （ct6, 61）. Results The morphologic of MSCs changed ob- viously and the immunohistochemical exam ination showed the positive staining of CK-19, integrin-α6, in- tegrin-β1 at 3rd and 6th day. The rate of positive cells were 9. 3%, 8.2%, 11.5% and 21.7%, 34. 1%, 39. 6%. The maker of CK-10 expressed negative. The cytokeratin-10 positive cells could not be observed. The number of positive ceils was decreased at 9rd day after training, the positive rate is 12. 2% , 18.6%, 16. 3%, at the same time, the CK-10 positive cells were increased and the positive rate was 10. 7%. Con- clusion The co-culture method of mesenchymal stem cells and HaCaT cell can make the MSCs differ enti- ate into epidermal cells in vitro.