HaCaT细胞诱导兔骨髓间充质干细胞表达角蛋白的初步研究

目的:探讨在体外培养的条件下HaCaT细胞诱导兔骨髓间充质干细胞(MSCs)表达角蛋白的可能性。方法:体外分离培养MSCs,流式细胞术检测CD34、CD45和CD44蛋白的表达以鉴定MSCs细胞。传代后与HaCaT共培养。分别在共培养3、7天后,密度梯度离心法分离MSCs和HaCaT,免疫细胞化学方法检测角蛋白Pan-cytokeratin(Pan-CK)、CK19的表达。结果:流式细胞术检测体外培养的MSCs CD34、CD45阴性,CD44阳性,表明MSCs培养成功。MSCs与HaCaT共培养3天、7天后,免疫细胞化学检测结果显示Pan-CK和CK19均为阳性。共培养7天的MSCs Pan-CK阳性率明显高于共培养3天的MSCs[分别为(29.1±8.3)%、(13.5±3.7)%,t=13.35,P<0.05],共培养7天的MSCs CK19阳性率与共培养3天的MSCs相比无统计学差异[分别为(3.3±0.8)%、(3.1±0.5)%,t=1.37,P>0.05]。结论:与HaCaT细胞共培养可诱导兔骨髓间充质干细胞向表皮细胞及表皮干细胞分化。

Investigation on HaCaT Cells Inducing Marrow Mesenchymal Stem Cells from Rabbit to Express Cytokeratins

Objective：To investigate the possibility of inducing marrow mesenchymal stem cells （ MSCs） from rabbit to express cytokeratin by co-culturing with HaCaT cells in vitro.Methods：MSCs were isolated and identified by detecting the expressions of CD 34,CD44,CD45 with flow cy-tometry,then the MSCs were co-cultured with HaCaT in vitro.After co-cultured for 3 and 7 days, the MSCs and HaCaT cell were separated , and Pan-cytokeratin and CK19 were detected by immu-nocytochemistry respectively .Results：Flow cytometry showed negative reaction in primary culture cells for CD34,CD45 and positive for CD44, which proved these were MSCs .After Co-cultured for 3 and 7 days, immunohistochemistry revealed that MSCs were all positive for Pan-cytokeratin （Pan-CK） and CK19.The positive rate of Pan-CK of the MSCs which co-cultured with HaCaT cell for 7 day as higher than that for 3 days,which was statistical significantly （29.1 ±8.3%, 13.5 ±3.7%, t=13.35, P0.05）.Conclusion：MSCs can express Pan-CK and CK19 by co-culturing with HaCaT cell , which indicated that MSCs might has the potential ac-tion to differentiate into epidermal stem cells .