体外培养3月龄和12月龄大鼠骨髓基质细胞增殖分化能力比较

The Difference in Proliferation and Differentiation Ability of Cultured Marrow Stromal Cells Between 3-Month-Old and 12-Month-Old Rats in Vitro

目的 对比研究 3月龄与 12月龄大鼠骨髓基质细胞 (r MSCs)增殖能力和成骨、成脂肪分化能力。方法 体外培养 3月龄、12月龄大鼠骨髓基质细胞 ,用血球计数板描绘生长曲线比较增殖能力 ;培养液中分别加成骨、成脂肪诱导剂 ,在不同时间行细胞化学染色 ,观察两组细胞成骨与成脂肪能力的变化。结果 12月龄r MSCs第 4代 (P4)及第 9代 (P9)较 3月龄 r MSCs同代细胞生长数量均下降。 12月龄 r MSCs P9较 P4生长数量下降。 3月龄 r MSCs P4、P9之间生长数量无显著差异。成骨诱导 1周后 ,12月龄 r MSCs对照与诱导组碱性磷酸酶(AL P)表达阳性率均低于 3月龄 r MSCs的同组细胞。成骨诱导 12天后 ,3月龄 r MSCs先于 12月龄 r MSCs组有钙化形成。成脂肪诱导第二天 12月龄 r MSCs先于 3月龄 r MSCs有脂滴生成。结论 12月龄 r MSCs生长能力和成骨能力较 3月龄 r MSCs降低 ,成脂肪能力较 3月龄 r

Objective To compare the difference in proliferation and differentiation (to osteoblast or adipocyte) ability of cultured rat marrow stromal cells (rMSCs) between 3-month-old and 12-month-old rats in vitro. Methods The rMSCs of 3-month-old and 12-month-old were cultured in vitro. The growth curves were depicted to compare their proliferation ability. Both 3-month-old and 12-month-old rMSCs were induced to osteoblasts or adipocytes by osteogenic inducer or adipogenic inducer. At different times, these cells were observed by histochemistry staining. Results The growing ability of both passage 4 (P4) and passage 9 (P9) of 12-month-old rMSCs decreased in comparison with that of 3-month-old rMSCs. The growing ability of P9 was less than that of P4 of 12-month-old rMSCs. The expression of alkaline phosphatase (ALP) of the control group and induced group of 12-month-old rMSCs was less than that of 3-month-old rMSCs after 1-week osteogenic induction. 12 days later, calcification was observed in 3-months-old group. Lipid droplets occurred in the cells of 12-month-old group after 2-day adipogenic induction, whereas the droplets occurred after 3-day or 4-day induction in the 3-month-old group. Conclusion The ability of proliferation and osteogenesis of 12-month-old group is weaker than that of 3-month-old group, but the ability of adipogenesis of 12-month-old group is stronger.