人胚胎干细胞向间充质干细胞分化的低血清诱导法的建立  

Establishment of a low-serum inducing method for the differentiation of human embryonic stem cells into mesenchymal stem cells

作　　者：冯年花[1] 曲学彬[2] 

机构地区：[1]首都医科大学附属北京朝阳医院医学研究中心,北京100020 [2]徐州医学院神经生物学教研室,江苏徐州221009

出　　处：《基础医学与临床》2016年第8期1130-1134,共5页Basic and Clinical Medicine

摘　　要：目的建立人胚胎干细胞向间充质干细胞分化的低血清诱导方法。方法用低血清培养体系培养人胚胎干细胞，每3d半量换液，7-10d后消化成单细胞用扩增培养基培养，每3d传代1次。结果人胚胎干细胞呈克隆样增殖，边缘呈锯齿状，表达多能干细胞标志OCT4、SOX2和NANOG。诱导3d后，克隆间隙出现体积较小的成纤维样细胞，增殖能力强，经两次传代后可获得形态一致的长梭形细胞。这些细胞表达间质细胞标志VIMENTIN，细胞均表达CD105、CD90、CD73、CD44和HLA-ABC，不表达CD34、CD45和HLA-DR，且具有成脂和成骨分化能力。结论用低血清诱导法能快速获得较高纯度的人胚胎干细胞源性间充质干细胞。Objective To establish a differentiation inducing method of mesenchymal stem cells （MSCs） from human embryonic stem cells （hESCs） using low-serum containing medium. Methods The induction was initia- ted by culturing human embryonic stem cells in low-serum containing medium, 7 ~ 10 days later, cells were con- fluent and dissociated into single cells then transfered into subculture. Results hESCs were cultured in feeder free condition and grew as clones with serrated edges, all clones expressed pluripoteney markes Oct4, Sox2 and Nanog. After 3 days＇induction, fibroblast-like cells with small volume appeared between hESCs clones. These cells grew as clusters and possessed high proliferation potential. Cells with uniform spindly morphology were ob- tained after 2 passage. All cells expressed mesenchymal marker-vimentin. Flowcytometry analysis showed that these cells expressed CD105, CD90, CD73, CD44 and HLA-ABC but not CD34, CD45 and HLA-DR. Under certain conditions, these cells may differentiate into osteoblasts and fat cells. Conclusions Human embryonic stem cell derived mesenchymal stem cells （hESC-MSCs） with high purity can be obtained in a short time by using low-serum containing induction medium.

关 键 词：间充质干细胞 胚胎干细胞 低血清诱导 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]