[1]肖宏涛),牛希华),刘毅)#.人脐带间充质干细胞的体外分离、培养及诱导分化*[J].郑州大学学报(医学版),2010,(02):213-216.
 XIAO Hongtao),NIU Xihua),LIU Yi).Isolation,culturation,and induced differentiation of human umbilical cord mesenchymal stem cells in vitro[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2010,(02):213-216.
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人脐带间充质干细胞的体外分离、培养及诱导分化*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2010年02期
页码:
213-216
栏目:
论著
出版日期:
2010-03-30

文章信息/Info

Title:
Isolation,culturation,and induced differentiation of human umbilical cord mesenchymal stem cells in vitro
作者:
肖宏涛1)牛希华1)刘毅2)#
1)郑州市第一人民医院河南省烧伤诊治网络中心郑州4500042)兰州军区兰州总医院烧伤整形科兰州730050
Author(s):
XIAO Hongtao1)NIU Xihua1)LIU Yi2)
1)Burn Diagnosis and Treatment Network Center in Henan Province,the First People’s Hospital of Zhengzhou,Zhengzhou 4500042)Department of Burn and Plastic Surgery, Lanzhou General Hospital,Lanzhou Command of PLA,Lanzhou 730050
关键词:
脐带间充质干细胞分离培养诱导分化体外
Keywords:
umbilical cord mesenchymal stem cellisolationculturationinduced differentiationin vitro
分类号:
Q813.1
文献标志码:
A
摘要:
体外分离、纯化及培养人脐带间充质干细胞(MSCs),并观察其生物学特性。方法:将剔除动静脉的新鲜人脐带组织切成小块培养,得到贴壁细胞,倒置显微镜观察细胞形态,绘制第1、5及10代细胞生长曲线;流式细胞仪测定细胞表面标记(CD13、CD44、CD14、CD34与HLADR);化学染色(油红O、茜素红染色)及RTPCR检测其体外诱导成脂和成骨分化的能力;RTPCR检测胚胎干细胞特异性标志基因OCT4。结果:体外培养4~6d后,有细胞从组织块中游出;细胞传代培养达10代后无明显的形态和增殖能力改变。培养细胞表达CD13与CD44,但CD14、CD34及HLADR呈阴性表达。体外诱导实验证实,该细胞具有成脂和成骨分化的能力。RTPCR显示其表达OCT4基因。结论:人脐带MSCs能在体外培养、扩增并且具有和骨髓MSCs类似的生物学特性,可作为组织工程种子细胞来源。
Abstract:
To explore the conditions of isolation, purification,and culture of human umbilical cord mesenchymal stem cells(MSCs) in vitro,and study their biological characteristics.Methods:After stripping off arteries and veins, the remaining parts of umbilical cord were cut into small sections and cultured. Adhere cells were obtained,and the morphology of the cells was observed under inverted phase contrast microscope. The growth curves of them were drawn and the surface antigens of human umbilical cord MSCs (CD13,CD44,CD14,CD34,and HLADR) were detected by flow cytometry.The potentiality of their osteogenic and adipogenic differentiation was detected by chemistry staining and RTPCR,and the stem cell marker gene OCT4 was detected by RTPCR.Results:Four to six days after primary culture,adhere cells came out of fragments and could be maintained for 10 passages without obvious changes in morphology or growth pattern. Flow cytometry analysis revealed that CD13 and CD44 were expressed on these cells’ surface,while CD14,CD34,and HLADR were negativeexpressed. The cells exhibited multipotential of differentiating into osteogenic and adipogenic cells.RTPCR results showed that these cells expressed OCT4.Conclusion:Human umbilical cord MSCs could be cultured and proliferated in vitro and have the similar biological morphology with bone marrow MSCs,and could be used for tissue engineering.

参考文献/References:

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30872689;全军“十一五”医学科学技术研究面上基金资助项目06MA079#通讯作者,男,1964年生,博士,教授,主任医师,研究方向:脂肪组织工程,Email:Liuzhih20002003@yahoo.com.cn
更新日期/Last Update: 2010-04-29