[1]叶发刚,夏长所,张卫兵,等.兔骨髓间充质干细胞的分离、培养与鉴定[J].青岛大学学报(医学版),2006,42(04):330-332.[doi:10.11712/qdyxy200604019]
 YE FA-GANG,XIA CHANG-SUO,ZHANG WEI-BING,et al.APPROACHES TO ISOLATE, CULTURE AND IDENTIFY THE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS OF RABBITS[J].JOURNAL OF QINGDAO UNIVERSITY (MEDICAL SCIENCES),2006,42(04):330-332.[doi:10.11712/qdyxy200604019]
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兔骨髓间充质干细胞的分离、培养与鉴定()
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《青岛大学学报(医学版)》[ISSN:2096-5532/CN:37-1217/R]

卷:
第42卷
期数:
2006年04期
页码:
330-332
栏目:
出版日期:
2006-12-25

文章信息/Info

Title:
APPROACHES TO ISOLATE, CULTURE AND IDENTIFY THE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS OF RABBITS
文章编号:
1672-4488(2006)04-0330-03
作者:
叶发刚1夏长所2张卫兵3夏仁云1
1 华中科技大学同济医学院附属同济医院骨科,湖北 武汉 430030; 2 青岛大学医学院附属医院创伤骨科; 3 南京鼓楼医院骨科
Author(s):
YE FA-GANG XIA CHANG-SUO ZHANG WEI-BING et al
Department of Orthopaedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
关键词:
骨髓间充质干细胞离心法梯密度 细胞培养技术
Keywords:
mesenchymal stem cells rabbits centrifugation density gradient cell culture techniques
分类号:
Q813.1+1
DOI:
10.11712/qdyxy200604019
文献标志码:
A
摘要:
目的 优化获取兔骨髓间充质干细胞并对其进行纯化、鉴定,同时对其生物学性状进行研究。方法取新生新西兰大耳白兔骨髓5.0 mL,采用密度梯度离心法、贴壁筛选法并结合传代对其进行纯化,观察细胞形态、超微结构和表面标志物的表达,从而对其进行鉴定。观察第1、3、5、8、10代细胞的增殖情况,并绘制出生长曲线。结果 经密度梯度离心、贴壁筛选并结合传代所得到的间充质干细胞很纯,第3代和第5代细胞形态单一,细胞排列具有典型的漩涡状结构。第1、3、5代细胞增殖能力强,生长旺盛;而第8、10代细胞增殖明显减弱。所分离培养的细胞表达CD44、CD90,不表达CD34,电镜观察为低分化细胞。结论 分离培养的细胞为间充质干细胞且成分单一,具有干细胞特性,第3、5代细胞很纯且其增殖能力强,适用于做以后的研究。
Abstract:
Objective  To explore the approaches to isolate,culture and identify the bone marrow-derived mesenchymal stem cells (BM-MSCs) of rabbits.
Methods  BM-MSCs were isolated from the bone marrow (5.0 mL) of the newborn rabbit via density gradient centrifugation. Light and electron microscopy were used to study the morphologic features and immunocytochemistry to examine the expression of cell surface antigens. The growth curve of cell proliferation was obtained based on the observation of the proliferation status of 1st, 3rd, 5th, 8th, and 10th generation.
Results  BM-MSCs of 1st, 3rd, 5th generation were very purified and all in the whirlpool-shape. They proliferated more rapidly than those of 8th and 10th generation. These cultured cells showed immunoreactivity to CD44 and CD90 but not CD34 and low-differentiation under the electron microscopy.
Conclusion Purified BM-MSCs can be obtained via this protocol. Cells of 1st , 3rd,5th generation with high proliferation are fit to the further experiment.
更新日期/Last Update: 2013-12-16