蒙古羊骨髓间充质干细胞的分离培养及多向诱导分化

中国畜牧兽医 ›› 2013, Vol. 40 ›› Issue (8): 7-12.

蒙古羊骨髓间充质干细胞的分离培养及多向诱导分化

韦林盖^1,2, 刘宗正^1,2, 高晋芳^1,2, 周欢敏^1,2, 张焱如^1,2

1. 内蒙古农业大学生命科学学院, 内蒙古呼和浩特 010018;
2. 内蒙古自治区生物制造重点实验室, 内蒙古呼和浩特 010018

收稿日期:2013-02-19 出版日期:2013-08-20 发布日期:2013-08-16
通讯作者: 张焱如 E-mail:yanru1964@163.com
作者简介:韦林盖(1986- ),男,海南人,硕士生,研究方向:动物成体干细胞。
基金资助:
内蒙古自然基金重大项目(2012ZD03);中国农大合作项目(2010)。

Isolation, Culture and Multiple Differentiation Induction of Mongolia Sheep Bone Marrow Mesenchymal Stem Cell

WEI Lin-gai^1,2, LIU Zong-zheng^1,2, GAO Jin-fang^1,2, ZHOU Huan-min^1,2, ZHANG Yan-ru^1,2

1. College of Life Sciences, Inner Mongolia Meteorological Institute, Hohhot 010018, China;
2. Key Laboratory of Inner Mongolia Autonomous Ragien Biobiomanufacture, Hohhot 010018, China

Received:2013-02-19 Online:2013-08-20 Published:2013-08-16

摘要/Abstract

摘要： 本研究旨在建立蒙古羊骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)体外分离、纯化、增殖方法,诱导其向脂肪细胞、成骨细胞及软骨细胞分化。穿刺法抽取蒙古羊骨髓组织,采用密度梯度离心法结合贴壁培养法分离纯化BMSC,增殖,并测定其生长曲线及细胞倍增时间。对第3代蒙古羊BMSC进行成脂、成骨及成软骨诱导,观察诱导后的细胞形态。采用油红O、茜素红、HE等染色法在组织学水平对BMSC进行成脂、成骨和成软骨分化鉴定。结果显示,蒙古羊BMSC呈现均一梭形成纤维细胞样生长,生长曲线呈S型,生长增殖能力良好。在不同分化诱导之后,细胞呈现脂肪细胞、成骨细胞及软骨细胞的表型特征。表明获得的蒙古羊BMSC具有多向分化潜能,所分离细胞确为骨髓间充质干细胞。

关键词: 蒙古羊; 骨髓间充质干细胞; 细胞培养; 多向分化

Abstract: To explore the optimal methods of isolating, puring, proliferating of Mongolia sheep bone marrow mesenchymal stem cells (BMSC), and their multiple differentiation potentiality.Bone marrow (BM) was collected from one 30-day-old sheep by puncture method.BMSC were harvested by gradient centrifuge and adhesive culture methods.Growth curves and cells double time of passaged BMSC were determined.The P3 BMSC were cultured in vitro under adipogenic,osteogenic and chondrogenic inductive environments and induced into adipocytes,osteoblates,chrondrocytes.Their adipogenic,osteogenic and chondrogenic differentiation properties were confirmed by histological staining such as oil red,Alizarin Red, HE.The Mongolia sheep BMSC were uniform spindle-shaped in appearance,showed active proliferative capacity and had S shape of growth curve. They could be induced to differentiate into adipocytes,osteoblasts and chondrocytes under different induction medium. The Mongolia sheep BMSC showed strong proliferation and multiple differentiation potential.It proved that the cells isolated were indeed the bone marrow mesenchymal stem cells.

Key words: Mongolia sheep; bone marrow mesenchymal stem cells (BMSC); cells culture; multiple differentiation

中图分类号:

S826.8⁺2

韦林盖, 刘宗正, 高晋芳, 周欢敏, 张焱如. 蒙古羊骨髓间充质干细胞的分离培养及多向诱导分化[J]. 中国畜牧兽医, 2013, 40(8): 7-12.

WEI Lin-gai, LIU Zong-zheng, GAO Jin-fang, ZHOU Huan-min, ZHANG Yan-ru. Isolation, Culture and Multiple Differentiation Induction of Mongolia Sheep Bone Marrow Mesenchymal Stem Cell[J]. , 2013, 40(8): 7-12.

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