中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8382-8385.doi: 10.3969/j.issn.1673-8225.2010.45.006

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

兔骨髓间充质干细胞的体外培养及多向诱导分化

张  蓉1，2，沈开金3，刘彦普2   

  1. 解放军兰州军区乌鲁木齐总医院，1颌面外科，3急诊科，新疆维吾尔自治区乌鲁木齐市   830000；2解放军第四军医大学口腔医院口腔颌面外科，陕西省西安市  710032
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 通讯作者: 刘彦普，博士，博士研究生导师，主任医师，解放军第四军医大学口腔医院口腔颌面外科，陕西省西安市 710032 jzkskj@sina.com
  • 作者简介:张蓉★，女，1971年生，河南省内乡县人，汉族，2003年解放军第四军医大学毕业，硕士，主治医师，主要从事口腔颌面外科方面的研究。zlaq2000@yahoo.com.cn

In vitro cultivation and multi-inducing differentiation of rabbit bone marrow mesenchymal stem cells

Zhang Rong 1,2, Shen Kai-jin3, Liu Yan-pu2   

  1. 1 Department of Maxillofacial Surgery, 3Emergency Department, Urumchi General Hospital of Lanzhou Military Area Command of Chinese PLA, Urumchi   830000, Xinjiang Uygur Autonomous Region, China;2 Department of Oral and Maxillofacial Surgery, Stomatological Hospital of Fourth Military Medical University of Chinese PLA, Xi’an   710032, Shaanxi Province, China
  • Online:2010-11-05 Published:2010-11-05
  • Contact: Liu Yan-pu, Doctor, Chief physician, Doctoral supervisor, Department of Oral and Maxillofacial Surgery, Stomatological Hospital of Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China
  • About author:Zhang Rong★, Master, Attending physician, Department of Maxillofacial Surgery, Urumchi General Hospital of Lanzhou Military Area Command of Chinese PLA, Urumchi 830000, Xinjiang Uygur Autonomous Region, China;Department of Oral and Maxillofacial Surgery, Stomatological Hospital of Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China zlaq2000@yahoo.com.cn

PDF

431

被引次数

19

摘要：

背景：骨髓间充质干细胞的分离获取及培养鉴定一直以来仍无统一标准和定论。
目的：体外获取兔骨髓间充质干细胞、纯化、扩增，同时研究其生物学特性及多向分化潜能。
方法：采用贴壁筛选法分离培养兔骨髓间充质干细胞，观察其增殖和生长特性，绘制生长曲线；倒置相差显微镜、苏木精-伊红染色及碱性磷酸酶染色观察细胞形态；并向成骨细胞、软骨细胞和血管内皮细胞多向诱导分化，分别采用钙结节茜素红染色、甲苯胺蓝染色和荆豆凝集素染色鉴定。
结果与结论：经贴壁筛选法得到的骨髓间充质干细胞性状稳定，为纺锤状，呈克隆样生长，碱性磷酸酶染色弱阳性；经成骨、成软骨、成血管内皮细胞诱导培养的骨髓间充质干细胞，表现出相应细胞的形态特征和生物学特征。提示，全骨髓培养法取材方便，骨髓间充质干细胞增殖能力强，在适宜条件下能够多向分化。

关键词: 间充质干细胞, 分离, 培养, 诱导分化, 生物学特性

Abstract:

BACKGROUND: There is still no unified standard and final conclusion of isolation, culture and evaluation of bone marrow mesenchymal stem cells (BMSCs).
OBJECTIVE: To observe the biological characteristics and multi-differentiation potential of BMSCs through in vitro cultivation, purification, and amplification.
METHODS: BMSCs were isolated and cultured by adherence screening method, followed by observation of proliferation and growth characteristics, for drawing growth curves; inverted phase contrast microscope, hemotoxylun and eosin staining, and alkaline phosphatase staining were used to observe the cell morphology; BMSCs were induced to differentiate into osteoblasts, chondrocytes and vascular endothelial cells, which were identified by calcified nodules-Alizarin red, toluidine blue and Ules europaeus agglutinin (UEA) staining respectively.
RESULTS AND CONCLUSION: By the method of adherence screening, BMSCs were found in stable characteristic, appeared morphologically in spindle and colony-shape, expressed weakly positive of Alkaline phosphatase staining. BMSCs presented with the corresponding cytomorphology and cytobiology characteristics after inducing cultivation of osteoblasts, chondrocytes and vascular endothelial cells. It revealed that adherence screening method is relatively simple and feasible. Furthermore, BMSCs have a good ability of proliferation and could multi-differenrate under suitable condition.

中图分类号: