增强型绿色荧光蛋白转基因小鼠脂肪间充质干细胞的多向分化潜能

Multilineage Differentiation Potential of Adipose-derived Mesenchymal Stem Cells from Enhanced Green Fluorescent Protein Transgenic Mouse

目的分离、培养增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)转基因小鼠脂肪间充质干细胞(adipose-derived mesenchymal stem cells,ADMSCs),并研究ADMSC在体外定向诱导下的多向分化潜能。方法分离EGFP转基因小鼠腹股沟的皮下脂肪组织,剪碎后胶原酶消化,含10%优等胎牛血清(fetal calf serum,FBS)的α-MEM培养液培养,取第3代EGFP-ADMSCs进行成骨、成脂诱导,并分别采用茜素红钙盐染色和油红O染色进行鉴定。结果成功获得了稳定表达EGFP的ADMSC;EGFP-ADMSCs经成骨诱导21 d后茜素红染色可见橘红色钙盐沉积,经成脂诱导14 d后油红O染色阳性,并且分化后细胞的EGFP表达不受影响。结论 EGFP转基因小鼠来源的脂肪间充质干细胞具有普通小鼠ADMSCs相同的自我更新增殖和多向分化的干细胞特性,并且能稳定表达EGFP,可以成为研究干细胞修复受损组织的作用机制的有效示踪工具。

Objective To isolate and culture adipose-derived mesenchymai stem ceils （ADMSC） of enhanced green fluorescent protein （EGFP） transgenic mice, and study the multilineage differentiation potential of ADMSC under different induction mediums. Methods Adipose tissue from the groin region of EGFP transgenic mice were sheared and digested in collagenase solution, and cultured in the medium with 10% fetal bovine serum and α-MEM. The passage 3 EGFP- ADM- SCs were induced into lipoblast and osteoblast, and identified by Alizarin red staining and oil red O staining. Results ADMSCs were obtained successfully from the EGFP transgenic mice. After 21 days of calcium induction,jacinth calcium salts could be detected in the cells by alizarin red staining. At day 14 after adipo induction, the cells showed positive oil red O staining,and the expression of EGFP protein did not change in differentiated cells. Conclusion ADMSCs harvested from EGFP transgenic mice have similar ability to self-renewal and similar multipotency in differentiation as those from cells of the ordinary mice, and stably express EGFP. The cells could be used as an effective tracking tool.