马骨髓间充质干细胞向软骨细胞的诱导分化

doi:10.16431/j.cnki.1671-7236.2017.02.017

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (2): 417-424.doi: 10.16431/j.cnki.1671-7236.2017.02.017

马骨髓间充质干细胞向软骨细胞的诱导分化

刘乐^1,2, 王彦妮^1,2, 庞玉娟^1,2, 张婷婷¹, 刘紫佳¹, 朱梦¹, 曹俊伟^1,2, 张焱如^1,2, 刘春霞^1,2

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

收稿日期:2016-07-29 出版日期:2017-02-20 发布日期:2017-02-25
通讯作者: 刘春霞 E-mail:lcx.imau@163.com
作者简介:刘乐(1990-), 男, 内蒙古乌兰察布人, 硕士生, 研究方向:动物成体干细胞, E-mail:coke1990@qq.com
基金资助:
内蒙古自治区自然科学基金（2016MS0315）；内蒙古自治区高等学校科学研究项目（NJZY16066）；内蒙古农业大学生命科学学院师生协同科技创新项目（201501）

Study on Inducing Equine Bone Marrow Mesenchymal Stem Cells Differentiated into Chondrocytes

LIU Le^1,2, WANG Yan-ni^1,2, PANG Yu-juan^1,2, ZHANG Ting-ting¹, LIU Zi-jia¹, ZHU Meng¹, CAO Jun-wei^1,2, ZHANG Yan-ru^1,2, LIU Chun-xia^1,2

1. College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Key Laboratory of Inner Mongolia Autonomous Region Biological Manufacturing, Hohhot 010018, China

Received:2016-07-29 Online:2017-02-20 Published:2017-02-25

摘要/Abstract

摘要：

试验旨在建立马骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）细胞系，并诱导其向软骨细胞分化。通过获取马BMSCs，进行细胞培养和纯化，对第3代（P3）细胞进行干细胞特性鉴定，并诱导其向软骨细胞分化，对分化后的细胞染色，并检测其软骨细胞特异性基因的表达。结果显示，获得的马BMSCs表达标记基因Sox2和Nanog，并表达间充质干细胞表面标记因子CD44、CD90和CD105，不表达造血细胞表面标志物CD34和CD45。P3代细胞经诱导培养后形态发生改变，阿尔新蓝染色为阳性，并表达软骨细胞特异基因Col，且其表达量随着诱导分化时间的增加而增高。综上表明，本试验建立了马BMSC细胞系，并成功诱导其分化为软骨细胞，为软骨损伤的干细胞治疗提供了试验依据。

关键词: 马; 骨髓间充质干细胞; 诱导分化; 软骨细胞

Abstract:

This study was aimed to establish equine bone marrow mesenchymal stem cells(BMSCs) line and induce it to differentiate into chondrocytes. The BMSCs were collected by cutting the bone and flushing the cutting surface by PBS, and then the bone marrow was washed with PBS,the collected cells were cultured after centrifugation. The cells were purified by passaging,the stem cell properties were tested before the induction. And the cells were also appraised by the expression of the special gene of chondrocytes as well as staining the differentiated cells with Alcian blue to insure the induction was effective. The obtained BMSCs expressed Sox2 and Nanog genes, which were the stem cell special genes, and also expressed CD44, CD90 and CD105 genes, but absent of CD34 and CD45 genes. The shapes of the 3rd passage BMSCs were changed after cultured in inducing medium for a few days. Furthermore, the cells were positive to Alcian blue staining, increased expression of the Col special gene of chondrocyte day by day as well. According to this study, the BMSC line was established, and the BMSCs were induced and differentiate into chondrocytes successfully.

Key words: equine; bone marrow mesenchymal stem cells; induced differentiation; chondrocyte

中图分类号:

Q254

刘乐, 王彦妮, 庞玉娟, 张婷婷, 刘紫佳, 朱梦, 曹俊伟, 张焱如, 刘春霞. 马骨髓间充质干细胞向软骨细胞的诱导分化[J]. 《中国畜牧兽医》, 2017, 44(2): 417-424.

LIU Le, WANG Yan-ni, PANG Yu-juan, ZHANG Ting-ting, LIU Zi-jia, ZHU Meng, CAO Jun-wei, ZHANG Yan-ru, LIU Chun-xia. Study on Inducing Equine Bone Marrow Mesenchymal Stem Cells Differentiated into Chondrocytes[J]. , 2017, 44(2): 417-424.

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马骨髓间充质干细胞向软骨细胞的诱导分化

Study on Inducing Equine Bone Marrow Mesenchymal Stem Cells Differentiated into Chondrocytes

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