周心涛,赵黎丙,闵新文,陈娇,郎明健
ZHOU Xintao,ZHAO Libing,MING Xinwen,CHEN Jiao,LANG Mingjian

• 1:湖北医药学院附属东风医院

摘要(Abstract)：

目的:探讨脐带血清(UCBS)分离、培养人胎盘间充质干细胞(PMSCs)的增殖及分化能力。方法:将UCBS处理后用于分离、培养PMSCs作为实验组,同体积FBS分离、培养PMSCs作为对照组,用相差显微镜观察实验组与对照组PMSCs的形态、流式细胞仪检测PMSCs表面标志、MTT法检测PMSCs的增殖能力。结果:10%UCBS的培养液分离纯化的PMSCs成梭形、增殖能力强,具有分化为成骨细胞和脂肪细胞的能力,高表达CD29、CD44及CD105,低表达或不表达CD34、CD45及HLA-DR;与对照组相比,UCBS组细胞增殖能力更强。结论:脐带血清能分离、培养及扩增PMSCs,培养的PMSCs的增殖能力及分化潜能好于FBS。
Objective: To investigate the proliferation and differentiation abilities of human placenta mesenchymal stem cells( PMSCs) isolated and cultured in human umbilical cord blood serum( UCBS). Methods: PMSCs isolated and cultured in UCBS served as experiment group and the same volume PMSCs isolated and cultured in FBS served as the control. PMSCs cell's morphology were observed under phase contrast microscope,flow cytometry were used to detected cell surface markers,MTT were used to test cell proliferation activity. Results: PMSCs isolated and cultured in 10% UCBS were spindle shaped,having high reproductive activity and could be induced to osteoblasts and adipocytes,highly expressed CD29,CD44 and CD105,but lowly expressed or didn't express CD34,CD45 and HLA-DR. The proliferation of experiment group was stronger than that of control group. Conclusions: UCBS can be used to isolate and culture PMSCs,and the proliferation and differentiation abilities of PMSCs in UCBS are better than those in FBS.

关键词(KeyWords)： 脐带;血清;间充质干细胞;流式细胞术;细胞培养
umbilical cord;serum;mesenchymal stem cell;flow cytometry;cell culture

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 周心涛,赵黎丙,闵新文,陈娇,郎明健
ZHOU Xintao,ZHAO Libing,MING Xinwen,CHEN Jiao,LANG Mingjian

DOI: 10.19367/j.cnki.1000-2707.2015.04.012

参考文献(References)：

• [1]张芬熙,洪艳,梁文妹.人脐带间充质干细胞的分离培养及超微结构特点研究[J].贵阳医学院学报,2013(1):5-9,15.
• [2]Montanucci P,Basta G,Pescara T,et al.New simple and rapid method for purification of mesenchymal stem cells from the human umbilical cord Wharton jelly[J].Tissue Eng Part A,2011(17):2651-2661.
• [3]姚旺祥.胎盘干细胞在组织工程中的应用进展[J].中国现代医生,2012(36):9-11.
• [4]Spees JL,Gregory CA,Singh H,et al.Internalized antigens must be removed to prepare Hypoimmunogenic mesenchymal stem cells for cell and gene therapy[J].Mol Ther,2004(5):747-756.
• [5]Tekkatte C,Gunasingh GP,Cherian KM,et al."Humanized"stem cell culture techniques:the animal serum controversy[J].Stem Cells Int,2011(2011):504-723.
• [6]Ang LP,Do TP,Thein ZM,et al.Ex vivo expansion of conjunctival and limbal epithelial cells using cord blood serum-supplemented culture medium[J].Invest Ophthalmol Vis Sci,2011(9):6138-6147.
• [7]Nimura A,Muneta T,Koga H,et al.Increased proliferation of human synovial mesenchymal stem cells with autologous human serum[J].Arthritis Rheum.2008(2):501-510.
• [8]Lin HT,Tarng YW,Chen YC,et al.Using human plasma supplemented medium to cultivate human bone marrow-derived mesenchymal stem cell and evaluation of its multiple-lineage Potential[J].Transplant Proc,2005(10):4504-4505.
• [9]Shetty P,Bharucha K,Tanavde V.Human umbilical cord blood serum can replace fetal bovine serum in the culture of mesenchymal stem cells[J].Cell Biol Int,2007(3):293-298.
• [10]Ma HY,Yao L,Yu YQ,et al.An effective and safe supplement for stem cells expansion ex vivo:cord blood serum[J].Cell Transplant,2012(5):857-869.
• [11]Ding Y,Yang H,Feng JB,et al.Human umbilical cordderived MSC culture:the replacement of animal sera with human cord blood plasma[J].In Vitro Cell Dev Biol Anim,2013(10):771-777.
• [12]刘玉磊,杨东生.银屑病患者皮肤间充质干细胞中表皮生长因子、转化生长因子β1的表达水平及其意义[J].实用临床医药杂志,2014(16):78-81.
• [13]邹金才.顺产产妇脐带血中抗氧化剂水平与分娩疼痛的相关性[J].贵阳医学院学报,2014(2):221-224.

文章评论(Comment)：