学术
影响脐血多能非造血成体干细胞分离的相关因素研究
李云涛 徐燕 孟恒星 葛薇 李桥川 万长春 于珍 李长虹 邱录贵
中国医学科学院中国协和医科大学、血液学研究所、血液病医院实验血液学国家重点实验室,天津300020
摘 要:
为了研究低血清条件下从脐血中分离多能非造血成体干细胞的相关影响因素，探索优化培养条件，通过不同培养液、接种密度、首次换液时间对脐血多能非造血成体干细胞生长的影响的比较，并以选定的条件对其培养扩增，进行免疫表型及诱导分化能力检测。结果表明：在低血清条件下，DMEM／F12培养液更适合于脐血多能非造血成体干细胞生长；1×10^6/cm^2是原代培养的适宜接种密度，原代第72小时首次换液为最佳换液时间。利用所选条件培养的细胞可在体外持续扩增，并具有良好的分化潜能。结论：建立了人脐血多能非造血成体干细胞的体外优化培养条件．为其在组织工程方面的应用作出了新的探索。[著者文摘]
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文章出处:
《中国实验血液学杂志》-2006年14卷4期 -731-736页
栏目信息:
分 类 号:
文献标识码:
A
文章编号:
1009-2137（2006）04-0731-06
Related Factors Affecting the Isolation of Multipotent Non-hema- topoietic Adult Stem Cells from Umbilical Cord Blood
LI Yun-Tao, XU Yan, MENG Hen-Xing, GE Wei, LI Qiao-Chuan, WAN Chang-Chun, YU Zhen, LI Chang-Hong, QIU Lu-Gui （State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical University, Tianjin 300020, China）
Abstract:
To investigate the related factors affecting the isolation of multipotent non-hematopoietic adult stem cells （MNASCs） from human umbilical cord blood in low serum （2%） condition, the isolation conditions were optimized and the yield of MNASCs was improved. MNASCs from human umbilical cord blood samples were isolated, and the effects of medium component, medium exchange time and initial plating density for isolation of MNASCs were studied. Then, the MNASCs were isolated and cultured in optimal condition, the surface antigen expression and differentiation potential of MNASCs were detected. The result showed that the medium of DMEM/F12 was better than IMDM and DMEM-LG for MNASCs culture in low serum condition, The optimal yield of MNASCs was obtained when mononuclear cells were cultured at a initial plating density of 1×10^6 cells/cm^2 and the medium was exchanged to remove the nonadherent cells after 72 hours of inoculation. MNASCs isolated and cultured under the above-mentioned conditions maintained a homogenous morphology, high potential ability of expansion and differentiation, It is concluded that culture conditions with low serum defined in this study is optimal for the successful isolation and expansion of umbilical cord blood MNASCs with high numbers for subsequent cellular therapeutic approaches.[著者文摘]
Key words:
stem cell; multipotent non-hematopoietic adult stem cell; umbilical cord blood; isolation efficiency
基金资助:
天津市科技发展计划项目,编号043803111
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