人羊膜间充质细胞分离培养及其干细胞特性研究

Study on isolation and culture of human amniotic mesenchymal cells and its stem cells characteristics

目的:研究人羊膜间充质细胞(Humanamnioticmesenchymalcells,HAMCs)的分离、培养及其干细胞特性,为羊膜间充质细胞在再生医学的潜在应用奠定实验基础。方法:无菌条件下取正常足月剖腹产胎儿的羊膜剪成碎片,经胰酶胶原酶序贯消化,DMEM/F12培养,倒置显微镜下观察其形态,MTT法检测其生长规律,免疫荧光的方法对细胞进行鉴定,定向诱导方法检测细胞的多向分化潜能。结果:来源于羊膜的间充质细胞,细胞免疫荧光显示SSEA-4,OCT-4阳性,具有很强的增殖能力,并且具有一定的多向分化能力,在特定条件下可分化为脂肪细胞和成骨细胞;结论:羊膜间充质细胞能够在体外分离、培养、扩增,并且具有干细胞特性。羊膜间充质细胞在再生医学和组织工程应用有很好的前景。

Objective：To investigate the isolation,culture and stem cells characteristics of human amniotic mesenchymal cells（HAMCs）for its potential application in regenerative medicine.Methods：Term placentas from healthy donor mothers obtained from caesarean sections were minced under sterile conditions.After sequential trypsin and collagenase digestion,cultivated in DMEM/F12,the morphololgy of HAMCs was observed under the microscope.Growth regularity was observed by MTT assay,the cell was identified by immunofluorescence,and it has multi-potential ability under specific conditions.Results：Mesenchymal cells derived from human amnion membrane,SSEA-4 and OCT-4 positive by immunofluorescence,It displays a strong proliferation and multipotential capacity.The cells can differentiate into fat cells and osteoblasts.Conclusions：HAMCs can isolation,culture and amplification in vitro,and these cells have stem cell properties.HAMCs has good prospects applications in regenerative medicine and tissue engineering.