龟板含药血清对大鼠骨髓间质干细胞体外增殖的影响

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。

Effects of Serum Containing Carapax et Plastrum Testudinis on In- Vitro Proliferation of Rat Mesenchymal Stem Cells

Objective] To observe the effect of serum containing Carapax et Plastrum Testudinis (CPT) on proliferation of rat mesenchymal stem cells (MSC). Methods] Rat mesenchymal stem cells dissociated from the bone marrow by density gradient method were cultured. After then, MSC were identified by marking of bromodeoxyuridine (Brdu) and staining of CD44. Effects of CPT - contained serum at different concentrations on the growth of MSC were observed by evaluating the morphological feature of MSC under light microscope,as well as MSC phenotypic characteristics after stained by immunohistochemical method, optical absorption value of MSC stained by thiazolyl blue colorimetry (MTT) and Brdu-positive cell counting after labeled by Brdu and proliferating cell nuclear antigen (PCNA). Results] CPT-contained serum at different concentrations could promote the proliferation of MSC and increase the number of Brdu positive and PCNA-positive cells in a dose-dependant manner, the difference being significant in comparison with the control group ( P < 0.05 or P < 0.01). Conclusion] CPT-contained serum can enhance the proliferation of MSC, and promoting the synthesis of deoxyribonucleic acid may be one of its kidney-reinforcing mechanisms.