CAJ | 학술논문

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(α6、β1)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、α6整合素、β1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、β1整合素、表达较前减少,阳性率为12.2％、18.6％、16.3％,CK1O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.
목적 탐토체외연합HaCaT세포공동배양유도골수간충질간세포(MSCs)향표피세포분화적가행성.방법 용취탄산지세포삽입판분층후연합공동배양HaCaT세포여MSCs,관찰배양3、6、9d후적세포형태,진행각단백(CK-19、CK-10)、정합소(α6、β1)염색병용류식세포의통계세포양성솔.결과 공동배양후세포형태변화명현,공배양3、6d후표피세포표지물CK-19、α6정합소、β1정합소면역형광염색양성,세포양성솔분별위9.3％、8.2％、11.5％화21.7％、34.1％、39.6％,CK10표체정음성；공배양9d후CK-19、α6정합소、β1정합소、표체교전감소,양성솔위12.2％、18.6％、16.3％,CK1O출현양성표체,세포양성솔위10.7％.결론 분층연합HaCaT세포공동배양가이유도골수간세포향표피세포진행분화.
Objective To explore the feasibility of the differentiation of bone mesenchymal stem cells (MSCs) into epidermal stem cells by the method of co-culture 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.Immunohistochemical staining and flow cytometry were carried out to detect the positive staining of cytokeratin (CK-19,CK-10) and integrin (α6,β1 ).Results The morphologic of MSCs changed obviously and the immunohistochemical exam ination showed the positive staining of CK-19,integrin-α6,integrin-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 cells 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％.Conclusion The co-culture method of mesenchymal stem cells and HaCaT cell can make the MSCs differ entiate into epidermal cells in vitro.