人脂肪基质细胞的分离、培养、增殖及传代稳定性

目的:建立人脂肪基质细胞分离、培养及扩增的方法,观察其增殖动力学行为,检测其传代稳定性。方法:通过吸脂术获取人的脂肪组织并分离脂肪基质细胞,在普通培养基中进行培养,观察细胞形态,绘制细胞增殖曲线,用油红O染色显示胞内脂滴生成,并检测其是否向脂肪细胞自动分化。结果:人的脂肪组织中可分离出基质细胞,在体外生长形态类似成纤维细胞;其增殖曲线呈S形。在原代及传代培养中均未见脂肪细胞或胞内脂滴生成。结论:成年人脂肪组织中存在的基质细胞可维持在未分化状态下稳定的增殖和传代。

Cultivation, proliferation, and passage of human adipose tissue-derived stromal cells in vitro

Objective:To investigate the optimal isolation,culturing conditions for human adipose tissue-derived stromal cells(hADSCs) and observe its proliferation patterns and stability of passage.Methods: Adipose tissues of healthy adults were obtained by liposuction.hADSCs were isolated from these adipose tissues and cultured in DMEM containing 10% FBS.The cells were observed under inverted microscope each day and cell growth was studied with cell counting.Intracellular lipid synthesis of hADSCs was examined by oil red staining and whether hADSCs would differentiate into adipocyte automatically was then identified.Results: hADSCs could be isolated from adult human adipose tissue,the cells were fibroblast-like and the growth curve of these cells was shown as "S"curve.No fat cell patterns were observed under microscope and no intracellular lipid synthesis was detected by oil red staining during primary culturing and passage of hADSCs.Conclusion: Adult adipose tissue is a confirmed source for hADSCs and the cells can be stably expanded and passaged as undifferentiated status in culture.