人脐带源间充质干细胞体外成脂、成骨、成软骨诱导分化  

In vitro study on differentiation of human umbilical cord mesenchymal stem cells into adipocytes,osteoblasts and chondrocytes

作　　者：汪兆艳[1] 杨印祥[1] 王倩[1] 王彩英[1] 屈素清[1] 金慧玉[1] 栾佐[1] 

机构地区：[1]海军总医院儿科,北京100048

出　　处：《转化医学杂志》2013年第6期329-331,335,共4页Translational Medicine Journal

摘　　要：目的 体外分离培养脐带来源间充质干细胞,确定其向脂肪、骨、软骨细胞的分化能力.方法 取新鲜的脐带组织,剔除脐动静脉,剩余部分剪成极为细小的组织块,0.2%Ⅱ型胶原酶37℃静止消化17 h,用含10%胎牛血清的达尔伯克改良伊格尔培养基/Ham＇s F12培养基终止消化,培养3 ~ 5 d后得到较为均一的贴壁细胞.取第3代细胞做流式检测,并定向诱导分化,14 d后成脂、成骨染色鉴定,21 d后取诱导分化的细胞团做冰冻切片,阿尔法蓝染色软骨鉴定.结果 体外培养的间充质干细胞高表达CD29、CD13、CD90、CD105、CD44等间充质细胞标志物,不表达CD34、CD45、人类白细胞抗原DR等造血细胞标志物;诱导分化后油红O染色、冯库萨染色均为阳性,冰冻切片阿尔法蓝染色呈明显阳性.结论 人脐带间充质干细胞在体外能够培养并定向分化为脂肪、骨、软骨细胞.Objective This article investigated the feasibility of inducing human umbilical mesenchymal stem cells （MSCs） into adipoeytes, osteoblasts and chondrocytes. Methods Taken fresh umbilical cord tissue to strip arteries and veins. The remaining parts of umbilical cord were cut into extremely small sections, digested with 0.2% collagenase for 17 hours at 37℃. Terminated di- gestion with DMEM/Ham＇s F12 medium containing 10% fetal bovine serum, then culturing for 3 to 5 days, we got adherent cells. After three passages, cells were detected by flow cytometry and in- duced directional differentiation. After 14 days, induced umbilical cord MSCs were identified by oil red O and von Kossa Staining. At 21 days, the induced cell mass were cut into frozen sections and identified by Alcian blue staining. Results Culturing cells expressed MSCs makers, such as CD13, CD90, CD105, CD44, CD29, and CD44 didn＇t express hematopoietic cell markers,such as CD34, CD45,and human leukocyte antigen-DR （HLA-DR）. After induced differentiation, Oil Red O, von Kossa and Alcian blue Staining were positive. Conclusion Human umbilical mesenchymal stem cells are able to be induced differentiation into adipocytes, osteoblasts and chondrocytes.

关 键 词：脐带 间充质干细胞 脂肪细胞 成骨细胞 软骨细胞 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] Q254[医药卫生—基础医学]