人骨髓MSC分离培养及诱导分化为脂肪细胞的实验研究

The study of human bone marrow mesenchymal stem cells separation and inducation to adipocyte

目的建立一种分离纯化,培养扩增人骨髓MSCs的方法,并探讨体外诱导骨髓间充质干细胞分化为脂肪细胞。方法密度梯度离心,贴壁培养和消化时间控制相结合,分离纯化人骨髓间充质干细胞,并用马血清诱导分化为脂肪细胞。结果MSCs原代培养呈均匀分布的集落样生长,呈梭形,细胞传代稳定,在体外连续传代培养9代,未发生形态学改变,无衰老征象;传代培养MSCs(P3)在20%马血清的L-DMEM培养液中分化为脂肪细胞。结论骨髓MSCs体外增殖和传代能力强,通过离体培养可使体内环境下低丰度的MSCs实现数量扩增;骨髓MSCs在体外可诱导分化为脂肪细胞。

Objective The aim of this study is to research on human bone marrow MSCs isolating,purified,amplified method and induce to adipocyte. Methods Isolated and purified MSCs from bone marrow by density gradiend centrifugation and adhering to the culture plastic.After successive subculture and amplification,Passage 3 were induced in 20%horse serum L-DMEM ,observed MSCs morphological change and differentiated character.Results The isolated MSCs was quite stable in L-DMEM containing 10% FBS and the growth curves of passage 1,3,5were auite similar,exhibiting a large expensive potential and the typical fibroblast-like morphology. Passage 3 differentiated to adipocyte in L-DMEM containing 20% horse serum ,Red O staining result was positive. Condusion A method for isolation and purification in vitro ofMSCs from bone marroe has been established. MSCs are easy to be isolated,purified and amplified in vitro.