骨康方对大鼠MSCs体外向成骨细胞分化和ALP的影响

目的:观察骨康方对大鼠骨髓基质细胞(marrow stromal cells,MSCs)体外增殖并向成骨细胞分化和其对碱性磷酸酶(ALP)活性的作用。方法:用流式细胞仪鉴定第3代MSCs表面抗原,采用不同浓度的含骨康方药血清联合诱导剂对大鼠MSCs定向诱导分化,观察MSCs诱导前后形态学变化,MTT法观察各组对MSCs的增殖作用并绘制细胞生长曲线,检测ALP活性。结果:CD34表达为阴性,CD44表达为阳性。原代MSCs及诱导后形态正常。细胞生长曲线示各组MSCs数量均随时间延长增加,联合使用高、中浓度骨康方药加诱导剂可显著提高MSCs增殖,并向成骨细胞增殖。联合使用高、中、低不同浓度骨康方药加诱导剂均可显著提高MSCs分化为成骨细胞的ALP活性。结论:骨康方能促进大鼠体外MSCs增殖,并向成骨细胞分化。且能提高成骨细胞的ALP活性,从而增强其成骨能力。

Effects of "GuKang" on Differentiation of Rat Marrow Stromal Cells to Osteoblasts in Vitro

Objective:To study the effects of blood-serum containing pharmacon of GuKang on the proliferation of bone marrow stromal cells and differentiation to osteoblasts in vitro. Methods:The surface antigens of the 3rd generation MSCs were identified by flow cytornetry. The rat bone marrow stromal cells were cultured in the media with varied concentrations of blood-serum containing pharmacon of GuKang and inductor, and the morphological changes were observed before and after being induced.MTT was used to observe the proliferation and the growth curve was obtained. The alkaline phosphata (ALP) activity was detected. Results:CD34 expression was negative,and CD44 expression was positive.The MSCs mot-phs of original generation and after induction were normal.The growth curve showed that the MSCs quantities of each group had increased accompanied with time.High concentration and middle concentrations of GuKang along with inductor significantly enhanced MSCs proliferation.High,middle and low concentrations of GuKang combined with inductor all significantly enhanced the ALP activity of MSCs induced to osteoblast. Conclusion:GuKang could significantly enhance the proliferation rat bone marrow stromal cells in vitro,and promote the ALP synthesis of MSCs,thus strengthen osteogene-sis.