人与家兔脂肪来源干细胞体外培养特性的比较

Comparison of ex vivo culture characteristics between human and rabbit adipose derived stem cells

背景:不同种属来源的脂肪来源干细胞在体外培养时特性是否存在差异目前尚未定论。目的:观察在相同培养条件下,人与家兔脂肪来源干细胞体外培养特性的异同。方法:体外分离人腹部取皮植皮术来源的脂肪来源干细胞、家兔背部皮下脂肪来源的脂肪来源干细胞,体外培养并传代,观察各自生长形态,取第3代脂肪来源干细胞,比较二者生长及增殖能力、表面CD分子鉴定情况及成脂、成骨分化能力。结果与结论:人和家兔皮下脂肪均能在体外分离出"成纤维细胞样"贴壁生长呈长梭形的细胞;人脂肪来源干细胞一般6～8d可传代,兔脂肪来源干细胞则需要四五天传代。四唑盐结果显示兔、人脂肪来源干细胞分别在第4,6天达到生长高峰。表面标记流式鉴定二者均显示CD29+CD31-。体外分离培养的人脂肪来源干细胞和兔脂肪来源干细胞均具有干细胞的培养特性。与人脂肪来源干细胞相比,兔脂肪来源干细胞具有更强的增殖和诱导成脂能力,但诱导成骨能力较差,家兔是做脂肪移植研究实验动物不错的选择。

BACKGROUND：Whether there are differences of different sources of adipose derived stem/stromal cells（ASCs） when cultured in vitro has been poorly understood.OBJECTIVE：To investigate the differences of ex vivo culture characteristics between human and rabbit ASCs under the same culture condition.METHODS：Human ASCs（hASCs） were isolated from intact fat of abdomen harvesting of skin grafts.Rabbit ASCs（rASCs） were derived from subcutaneous fat tissue of back.The hASCs and rASCs were cultured and passaged in vitro and the morphology of the cells was observed.Passage 3 ASCs were used to compare the ability of growth and proliferation,identification of CD surface molecules and the ability of adipogenic and osteogenic differentiation.RESULTS AND CONCLUSION：Fibroblast-like adherent spindle-shaped cells could be isolated from both human and rabbit subcutaneous fat tissue.hASCs could passage after cultured for about 6-8 days,but rASCs were about 4-5 days.MTT results manifested that the growth climax of hASCs was the 4 th day,whereas rASCs was the 6 th day.Flow cytometry analysis showed the expression of CD29 ＋ CD31-could be seen both in human and rabbit ASCs.Both hASCs and rASCs had the cultural characteristics of stem cells.Compared with hASCs,rASCs had more powerful ability of proliferation and induction to fat whereas the ability of induction to bone was poor.Rabbit was a good choice of laboratory animal for fat transplantation research.