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[目的]观察益肾中药龟板血清对大鼠骨髓间质干细胞(mesenchymal stem cells,MSC)体外增殖的影响。[方法]使用密度梯度法分离大鼠骨髓间质干细胞进行培养,经5-溴脱氧尿嘧啶(Brdu)标记和CD44染色及两者双重染色鉴定后,分别从光学显微镜下的形态学特征、免疫细胞化学染色后的MSC表型特征、四甲基偶氯唑盐(MTT)染色后MSC的吸光值以及以Brdu、增殖细胞核抗原(PCNA)免疫组化染色后MSC的阳性细胞数变化等角度观察在培养液中添加不同浓度龟板血清条件下MSC的生长变化。[结果]不同浓度的龟板血清均以浓度依赖方式促进MSC增殖活力和增加Brdu、PCNA阳性细胞数,与对照组比较具有显著性差异(P<0.05或P<0.01)。[结论]龟板血清可促进MSC的增殖而有利于细胞脱氧核糖核酸(DNA)合成,推测这可能是龟板补肾的机理之一。
[Objective] To observe the effect of Yishen traditional Chinese medicine turtle shell serum on proliferation of rat mesenchymal stem cells (MSCs) in vitro. [Methods] Rat bone marrow mesenchymal stem cells were isolated and cultured by density gradient method. After identification by Brdu labeling and CD44 staining and their double staining, their morphological characteristics and immunity were respectively analyzed by light microscopy. The phenotypic characteristics of MSCs after cytochemical staining, the absorbance of MSCs after staining with Methyltetrazolium (MTT), and the change of MSC positive cells after immunohistochemical staining with Brdu and proliferating cell nuclear antigen (PCNA) The growth of MSCs was changed in the culture medium with different concentrations of turtle serum. [Results] The concentrations of turtle shell serum in different concentrations all promoted the proliferation of MSCs and increased the number of Brdu and PCNA positive cells in a concentration-dependent manner, which was significantly different from the control group (P<0.05 or P<0.01). [Conclusion] The turtle shell serum can promote the proliferation of MSCs, which is beneficial to the synthesis of cell DNA. It is speculated that this may be one of the mechanisms of kidney tortoise.