贴壁法培养不同龄小鼠骨髓间充质干细胞的生物学特点

Biological characteristics of marrow mesenchymal stem cells of mice of different ages cultured in vitro with adherent method

目的观察不同龄小鼠骨髓间充质干细胞在体外分离、培养、扩增的过程,并对其生物学特性进行初步比较分析,为下一步实验提供依据。方法选取出生1周、8周、16周的健康昆明小鼠,分成新生、青年、成年三组,采用贴壁法分离培养骨髓间充质干细胞,进行细胞形态学和贴壁率观察,绘制生长曲线。结果各组细胞约在9-12天80%-90%长满培养瓶底,融合成单层,传代后的骨髓间充质干细胞形态更加单一,为排列更加有序的成纤维细胞样细胞。贴壁率的观察和生长曲线显示,在相同的培养条件下,新生小鼠细胞增殖最快,细胞可扩增能力最强,而成年小鼠细胞增殖最慢,细胞可扩增能力最弱,青年小鼠细胞介于两者之间。结论利用贴壁法可以从不同龄小鼠骨髓中分离出间充质干细胞,并可在体外传代扩增,干细胞的活性和增殖能力随着年龄的增加而降低。

Objecttve To observe the feature of extraction,separation, culture and passage of marrow mesenchymal stem cells from mice at different ages in vitro and investigate the biological characteristics of MSCs for provide experimental background for further study. Methods Kunming mice aged 1 week, 8 weeks, 16 weeks were used to isolate MSCs. Marrow mesenchhymal stem cells with adherence were cultured, passaged, amplifled and purified in vitro. Living eharactexistics and adhesive rate of the primary and generations were observed respectively. Growth curve was drawn. Remits MSCs were intact and fibroblast cell-shaped. The growth curve and adhesive rate showed that eeUs from newborn mice grew faster than those from young and adult mice at the same condition. Conclusion Different age mice＇s MSCs can be isohted and amplified with adherent method in vitro. The MSCs from younger mice showed a stronger capability of growth and proliferation compared with the MSCs from the older ones.