两种不同方法诱导脂肪干细胞向表皮细胞分化效果的比较

Differentiation of adipose stem cells into epidermal cells induced by two methods

目的比较两种不同方法培养诱导脂肪干细胞（ASCs）向表皮细胞分化效果的差异，以寻找更好的诱导分化方法。方法利用Transwell装置共培养HaCaT细胞与ASCs为共同培养组；在ASCs培养液中加入表皮生长因子（EGF）进行诱导培养为EGF诱导组；两种方法培养3、6d后的ASCs通过进行表皮细胞标志物角蛋白-19（CK-19）、B1整合素和广谱细胞角蛋白Pan—cytokeratin（Pan—CK）染色检测并计数。结果共培养法诱导3d后的ASCs细胞CK-19、B1整合素、Pan—CK阳性率分别为（25．8±4．1）％、（29．2±3．9）％、（18．3±2．7）％，明显高于EGF诱导组的细胞阳性率，差异有统计学意义（P〈0．05）。共培养法诱导6d后的ASCs细胞标记物CK-19、B1整合素、Pan—CK阳性率分别为（34．1±5．7）％、（42．8±4．3）％、（29．4±3．3）％，显著高于EGF诱导组的细胞阳性率（P〈0．01）。结果提示采用共同培养法获得的表皮细胞数目多于EGF诱导组，差异有统计学意义（P〈0．05）。结论与HaCaT细胞共同培养诱导比单纯应用EGF诱导能够更好地促进ASCs向表皮细胞的转化。

Objective To find an optimal way to induce differentiation of adipose-derived stem cells （ASCs） into epidermal cells. Methods In co-culture group, HaCaT cells and ASCs were co-cultured by using Transwell apparatus. In epidermal growth factor （EGF） induction group, EGF was added into the culture medium of ASCs. After culture for 3 and 6 days, ASCs were detected by counting the number and dying the markers of epidermal cells such as cytokeratin 19 （ CK-19 ） , integrin 131 and pan-cytokeratin （Pan-CK）. Results The positive rate of CK-19, integrin 131 and Pan-CK in ASCs was （25.8 ± 4. 1 ）% , （29. 2 ±3.9）% and （ 18.3 ±2. 7）% respectively at 3rd day after culture by co-culture method, which was significantly higher than in EGF induction group （P 〈 0. 05）. The positive rate of CK-19, integrin 131 and Pan-CK in ASCs was （ 34. 1± 5.7 ） % , （42. 8 ±4. 3 ） % and （29.4 ±3.3 ） % respectively at 6th day after culture by co-culture method, which was significantly higher than in EGF induction group （P 〈 0. 01 ）. The number of epidermal ceils obtained by co-culture method was significantly more than that in EGF induction group. Conclusion Co-culture of ASCs with HaCaT cells is an optimal method to promote the differentiation of ASCs into epidermal ceils.