中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (14): 2550-2554.doi: 10.3969/j.issn.1673-8225.2012.14.016

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

不同细胞接种浓度影响人脐带间充质干细胞生长及多向分化

李  辛，刁  克，张凤香，孙大鹏   

  1. 辽宁医学院附属第一医院心外科，辽宁省锦州市 121001
  • 收稿日期:2012-01-09 修回日期:2012-02-20 出版日期:2012-04-01 发布日期:2012-04-01
  • 通讯作者: 张凤香，博士，主治医师，辽宁医学院附属第一医院心外科，辽宁省锦州市 121001 zhangfengxiang64@163.com
  • 作者简介:李辛，女，1979年生，辽宁省锦州市人，汉族，2010年辽宁医学院毕业，主要从事干细胞的研究。cctv138416@163.com

Effects of different implantation densities on the growth and multi-directional differentiation of human umbilical cord mesenchymal stem cells 

Li Xin, Diao Ke, Zhang Feng-xiang, Sun Da-peng   

  1. Department of Cardiac Surgery, First Affiliated Hospital of Liaoning Medical University, Jinzhou  121001, Liaoning Province, China
  • Received:2012-01-09 Revised:2012-02-20 Online:2012-04-01 Published:2012-04-01
  • Contact: author: Zhang Feng-xiang, M.D., Attending physician, Department of Cardiac Surgery, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning Province, China zhangfengxiang64@163.com
  • About author:Li Xin, Department of Cardiac Surgery, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning Province, China cctv138416@163.com

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364

被引次数

2

摘要：

背景：在干细胞培养过程中寻找合理的细胞接种浓度，从而得到能稳定、高效生长的干细胞非常重要。
目的：观察不同接种浓度对人脐带间充质干细胞原代培养及分化为神经元样、胰岛样、脂肪样细胞的影响。
方法：分离人脐静脉内皮及内皮下层细胞，按不同接种浓度分组进行原代培养，记录原代培养时间并进行传代。取第1代人脐带间充质干细胞，体外诱导其向神经元样、胰岛样、脂肪样细胞分化。
结果与结论：原代培养发现以5×105~1×106/cm2细胞浓度接种的人脐带间充质干细胞生长状态最佳。经β-巯基乙醇诱导6 h，细胞即表达Nestin、神经丝蛋白及胶质纤维酸性蛋白；经尼克酰胺，活化素A，胰高血糖素样肽1诱导21 d可见明显的胰岛样细胞团，双硫腙染色呈砖红色；经成脂诱导培养基诱导后细胞油红O染色阳性，有明显脂滴出现。可见选取合适的细胞浓度可以快速获得大量的人脐带间充质干细胞，且该细胞经体外诱导可向神经元样、胰岛样及脂肪样细胞分化。
关键词：细胞浓度；脐带间充质干细胞；原代培养；分化；神经元样细胞；胰岛样细胞；脂肪样细胞
缩略语注释：MSCs：mesenchymal stem cells，间充质干细胞
doi:10.3969/j.issn.1673-8225.2012.14.016

关键词: 细胞浓度, 脐带间充质干细胞, 原代培养, 分化, 神经元样细胞, 胰岛样细胞, 脂肪样细胞

Abstract:

BACKGROUND: It is important to search a rational cell implantation density to achieve stably and high efficiently growing stem cells during the culture of stem cells.
OBJECTIVE: To investigate the effects of different implantation densities on primary culture of human umbilical cord mesenchymal stem cells and their differentiation into neuron-, islet-, and adipose-like cells.
METHODS: Human umbilical cord venous endothelial and subendothelial layer cells were isolated and grouped according to different implantation densities for primary culture. The time for primary culture was recorded and cell passage was performed. Passage 1 human umbilical cord mesenchymal stem cells were in vitro induced to differentiate into neuron-, islet-, and adipose-like cells.
RESULTS AND CONCLUSION: Human umbilical cord mensenchymal stem cells at an implantation density of 5×105-1 ×106/cm2 cell density showed optimal growth. After induced by β-sulfhydryl alcohol for 6 hours, human umbilical cord mesenchymal stem cells expressed nestin, neurofilament protein and glial fibrillary acidic protein. After induced by niacinamide, activin A, glucagon-like peptide 1 for 21 days, obvious islet-like cell masses appeared and were stained red by dithizone. After induced by adipogenic culture medium, cells were positive for oil red O staining, with appearance of obvious liquid drops. A large number of human umbilical cord mesenchymal stem cells could be quickly harvested at a proper implantation density and the cells could be induced to differentiate into neuron-, islet-, and adipose-like cells.

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