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目的评估体外分离培养皮肤间充质干细胞(sMSCs)的适宜方法。方法采用低血清培养和无血清培养两种不同的原代培养法,体外培养皮肤间充质干细胞,通过观察细胞形态特征及流式细胞技术进行鉴定,比较其差异。结果两种培养法均能培养出形态均一、成熟的皮肤间充质干细胞,但是低血清法培养细胞所用时间8.93d明显小于无血清培养法所需时间15.93d,差异有统计学意义(P<0.001),且低血清法培养出的细胞较无血清培养方法得到的皮肤间充质干细胞的CD44阳性率较高,CD45阴性率较低,差异有统计学意义(P<0.001)。结论低血清培养法是一种适用于临床试验研究皮肤间充质干细胞的原代培养方法。
Objective To evaluate the appropriate method for the isolation and culture of human dermal mesenchymal stem cells (sMSCs) in vitro. Methods Two kinds of primary culture methods of low serum culture and serum-free culture were used to culture skin mesenchymal stem cells in vitro. The morphological characteristics and flow cytometry were used to identify and compare the differences. Results Both of the two culture methods could produce uniform and mature skin mesenchymal stem cells. However, the time for culture of cells in low serum was 8.93 days, which was significantly shorter than that of serum - free culture for 15.93 days (P < 0.001). The CD44 positive rate and CD45 negative rate of skin mesenchymal stem cells obtained by low serum culture method were higher than those without serum culture method, the difference was statistically significant (P <0.001). Conclusion Low serum culture method is a suitable primary culture method for clinical trials of mesenchymal stem cells.