[1]王萧枫,童培建,金红婷,等.兔骨髓间充质干细胞的体外培养及相关检测的实验研究[J].中医正骨,2011,23(11):3-6.
　WANG Xiao-feng*,TONG Pei-Jian,JIN Hong-ting,et al.Approaches To Isolate,Culture and Identify The Rabbit Marrow Mesenchymal Stem Cells[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2011,23(11):3-6.
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兔骨髓间充质干细胞的体外培养及相关检测的实验研究()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第23卷
期数:
2011年11期
页码:
3-6
栏目:
基础研究
出版日期:
2011-11-30

文章信息/Info

Title:
Approaches To Isolate,Culture and Identify The Rabbit Marrow Mesenchymal Stem Cells
作者:
王萧枫1; 童培建2; 金红婷2; 吴银生1
1.浙江省温州市中西医结合医院,浙江 温州 325000; 2.浙江中医药大学,浙江 杭州 310053
Author(s):
WANG Xiao-feng*; TONG Pei-Jian; JIN Hong-ting; et al.*
Hospital of the Integration of Traditional Chinese and Western Medicine of Wenzhou City,Wenzhou 325000,Zhejiang,China
关键词:
间充质干细胞 体外培养 诱导分化 流式细胞术
Keywords:
Mesenchymal stem cells;  Cell culture techniques
摘要:
目的:建立体外培养扩增、检测鉴定兔骨髓间充质干细胞的方法,观察其生物学特性。方法:采用密度梯度离心法和贴壁分离法相结合,体外培养扩增兔骨髓间充质干细胞,观察细胞生长速度和形态变化; 选择第3代细胞分别以特定培养液诱导,观察其向成骨细胞、成脂细胞、成软骨细胞的分化能力,采用流式细胞学方法分别检测CD29、CD44、CD34、CD45的表达情况。结果:培养的细胞长梭形,呈成纤维细胞样生长,具有塑料贴附性、传代细胞生长迅速,在诱导下均能分化为成熟的成骨细胞、软骨细胞和脂肪细胞。流式结果表明,CD29阳性细胞比率92.5%,CD44阳性细胞比率86%; CD34阳性比率0.77%,CD45阳性细胞比率0.32%。结论:应用密度梯度分离法和贴壁筛选法相结合能从兔骨髓中分离培养出高纯度的兔MSCs,生长旺盛、增殖能力强,在条件诱导下具有向成骨细胞、脂肪细胞和软骨细胞分化的潜能。
Abstract:
Objective:To explore the approaches to isolate,culture and identify the bone marrow mesenchymal stem cells(MSCs)of rabbits in vitro.Methods:Rabbit MSCs were extracted through the density gradient centrifugation and separation of the combination adherent,cell growth rate and morphological changes was observed using the inverted microscope,select the P3 cells induced using special media supplemented,observe the feasibility of differentiation into bone cells,fat cells,cartilage cells.Expression of CD29,CD44,CD34,CD45 was detected using flow cytometry.Results:The cultured cells showing long fusiform and fibroblast-like growth,plastic attachment and rapid growth of passage cell,they can differentiate into mature bone cells,cartilage cells and fat cells under induction.Flow cytometry results showed that for the P3 cells,about 92.5% of the cells expressed CD29,about 86% of the cells expressed CD44,0.77% of the cells expressed CD34,0.32% of the cells expressed CD45.Conclusion:The density gradient separation and screening adherence combination isolated from rabbit bone marrow in vitro with high purity MSCs,vigorous growth,proliferation ability,under the conditions with the induction of osteoblasts,adipose cells and cartilage cells potential.

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更新日期/Last Update: 1900-01-01