文档介绍：脐血间充质干细胞分离、培养与鉴定
李启明1 程天民1 李宁2粟永萍1 冉新泽1
(,创伤、烧伤与复合伤国家重点实验室
, 高压氧中心重庆 400038)
摘要:目的研究脐带血(HUCB)中含有的间允质干细胞(MSCs)体外分离、培养条件,探讨其作为种子细
胞用于下一步实验研究的稳定性与实用性。方法无菌条件下取正常足生产的脐带血,经 CPDA-1 复合抗凝
剂抗凝,然后经过去红细胞处理,再用淋巴细胞分离液分离脐血的单个核细胞,DMEM/F12 培养基进行培养
和纯化获得贴壁细胞层,采用定向诱导分化方法检测鉴定获得的 MSCs 结果来源于脐血的单个核细胞经体
外培养贴壁后出现形态学上可见间充质样的细胞,问充质样细胞为类似成纤维样的细胞形态,但偏向半月
形弯曲状,经过成肌、成脂肪、成神经细胞诱导分化成功后确定为间充质干细胞。结论脐血来源的间充质
干细胞能够在体外分离、培养出有用的进一步实验所需要的种子细胞。
关键词:脐带血;间充质干细胞;细胞培养;诱导分化鉴定
中图法分类号: 文献标识码:
THE ISOLATION CULTURE AND IDENTIFICATION OF HUCA MSCs
LI Qi-ming, CHEN Tian-min LI ning, SU Yong-ping, RUAN xin-ze
( Key Laboratory of Trauma, Burns bined Injury, Institute bined Injury,
2. Hyperbaric oxygen Center Xinqiao Hospital,
Third Military Medical University, Chongqing, 400038, China)
Abstract: Objective To investigate the isolation , cultivation, differentiation and
identification of human umbilical cord blood(HUCB) mesenchymal stem cells(MSCs) in
vitro , and to observe the feasibility of using MSCs as seed cells in experimental
furthermore. Methods HUCB were collected from full term childbirth scheduled for Cesarean
section and vagina deliveries, a11 specimens was obtained sterilely with Preservative-free CPDA-1
compound natrium citricum. After lyse red blood cell, the cord blood mononuclear cell was
isolated by human lymphocyte separ