体外分离兔脂肪间充质干细胞多向诱导分化及鉴定的实验研究

Experimental study on multipotent and identification of rabbit adipose-derived stem cells isolated in vitro

目的研究兔脂肪间充质干细胞在体外多向诱导分化的潜力。方法对兔脂肪间充质干细胞体外分离培养,采用细胞活性检测(CCK-8)测生长曲线,流式细胞术鉴定细胞表型,兔脂肪间充质干细胞成脂、成骨、成软骨诱导分化。结果脂肪间充质干细胞经传代后可优化,P3培养第3天进入增殖期,细胞表型CD90呈阳性,CD34、CD11b呈阴性,贴壁细胞在第7天诱导形成脂肪小滴、骨结节,21 d Pellet结构的细胞团块可在TGF-β1的外源性刺激下形成II型胶原。结论脂肪间充质干细胞具有体外诱导成脂、成骨、成软骨分化的能力。

Objective To study the multipotent of rabbit adipose-derived stem cells to differentiate in vitro. Methods Rabbit adipose-derived stem cells were isolated and cultured in vitro. Cell counting kit(CCK-8) was used to measure growth curve and flow cytometry to identify cell phenotype, and adipogenic, osteogenic and chondrogenic differentiation of rabbit adipose-derived stem cells. Results Adipose-derived stem cells could be optimized after P3 cultures entered the proliferative phase on the 3 rd day. The cell phenotype was positive for CD90, while CD34 and CD11 b were negative, and adherent cells induced fat droplets and bone nodules on the 7 th day. The Pellet structure can form type II collagen under the culture with exogenous stimulation of TGF-β1. Conclusion Adipose-derived stem cells have the ability to induce adipogenic, osteogenic and chondrogenic differentiation in vitro.