年龄因素对大鼠脂肪干细胞成骨诱导分化能力的影响

Influence of Age on Osteogenic Differentiation of Rat Adipose-derived Stem Cells

目的探讨年龄因素对脂肪干细胞成骨分化能力的影响。方法通过密度梯度离心法分离不同年龄段ADSCs进行培养及传代,观察细胞生长情况,细胞培养至第3代进行诱导分化,各年龄组细胞均在成骨诱导分化后进行检测。观察其形态变化、细胞凋亡率、碱性磷酸酶活性及钙离子浓度测定。结果体外培养的ADSCs呈梭形,诱导条件下,各年龄组细胞形态由梭形变为立方形、多角形;经检测,随年龄的增长细胞的凋亡率在不断增大,幼年组细胞碱性磷酸酶活性较老年组高,钙离子浓度随年龄增加而降低。结论ADSCs成骨分化能力随着年龄的增加而降低。

Objective To investigate influence of ages on different osteogenic differentiation capability of rat adipose - derived stem cells （ADSCs） . Methods ADSCs were isolated by using density gradient centrifuge method of different ages, and cultured in vitro. The adhesive cells were preserved to passage, the third generation of cell culture was for induction differentiation. All groups of cells were cultured with DMEM medium, which contains 0. 1μmol/L Dexamethasone, 10 mmol/L β- glyeerophosphate and 50 μmol/ L Aseorbic Acid. The morphology change, alkaline phosphatase activity and calcium ion assaying were observed. Results The results showed that in vitro ADSCs showed fusiform. The passage ceils had high ADSCs proportion. Induced conditions morphology changed from fusiform into cubic shape and polygon, after the culture in the differentiation induction medium, alkaline phosphatase activity and calcium ion decreased with age. Conclusions Results indicate osteogenic differentiation capability of ADSCs decreases with aging. These findings have significant implications for the proposed clinical application of ADSCs.