小鼠脂肪源干细胞分离培养和成骨诱导前后造血调控因子的表达

背景：小鼠脂肪源干细胞的体外分离培养和诱导分化以及成骨诱导前后造血调控因子表达的报道甚少。目的：观察小鼠脂肪源干细胞的体外分离培养方法以及成骨诱导前后造血调控因子分子水平的表达。方法：采用酶消化法获得雄性C57BL/6小鼠脂肪来源的细胞,传代培养至第3代,分别进行成脂与成骨细胞诱导。结果与结论：小鼠脂肪来源的细胞体外培养呈梭形贴壁生长,可稳定传代。细胞表面标志CD29表达率强阳性,CD34表达率较低,CD106、CD49d低表达。成脂及成骨细胞诱导均为阳性。造血调控因子β-连环蛋白表达量最高,基质细胞衍生因子1和神经钙黏素次之,其余干细胞因子、巨噬细胞炎性蛋白1α、配体jagged1、粒细胞-巨噬细胞集落刺激因子均为低表达。结果证实,小鼠脂肪来源的细胞为脂肪源干细胞,其成骨诱导前后的造血调控因子均有表达,但差异无显著意义。

Isolation and culture of mouse adipose-derived stem cells and the expression of hematopoietic modulation factors before and after osteogenic induction

BACKGROUND： There are few reports about the isolation and culture of mouse adipose-derived stem cells in vitro and the expression of hematopoietic modulation factors before and after osteogenic induction. OBJECTIVE： To investigate the isolation and culture of mouse adipose-derived stem cells （ADSCs） and molecule level of hematopoietic modulation factors before and after osteogenic induction in vitro. METHODS： ADSCs were isolated from male C57BL/6 mouse by digesting adipose tissue with collagen typeⅠ, and expanded to three passages after primary culture in a control medium. Then the cells underwent the adipogenic and osteogenic induction. RESULTS AND CONCLUSION： ADSCs grew as spindle-shaped adherent cells when cultured in vitro, and could stably proliferate and be passed. The cells’ surface antigen phenotype was analyzed by flow cytometry, and the expression of CD29 was strongly positive, CD34 was lower, the expression of CD106 and CD49d was negative. ADSCs incubated in the adipogenic medium and osteogenic medium were stained positively. The expression of hematopoietic modulation factors-catenin was the highest, the stromal cell derived factor-1 and N-cadherin was followed by, the expression of the other factors such as stem cell factor, macrophage inflammatory protein-1α, ligand jagged1 and granulocyte macrophage colony-stimulating factor were all lower. It indicates that the cells from adipose tissue are ADSCs, and the expression of hematopoietic modulation factors is found before and after osteogenic induction, but has no significant difference.