A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone

被引:426
作者
Zhu, Heng [1 ]
Guo, Zi-Kuan [2 ]
Jiang, Xiao-Xia [1 ]
Li, Hong [1 ]
Wang, Xiao-Yan [1 ]
Yao, Hui-Yu [1 ]
Zhang, Yi [1 ]
Mao, Ning [1 ]
机构
[1] Chinese Acad Med Sci, Inst Basic Med Sci, Dept Cell Biol, Beijing 100730, Peoples R China
[2] Inst Radiat Med, Dept Expt Hematol, Beijing, Peoples R China
关键词
MARROW STROMAL CELLS; PROGENITOR CELLS; TRABECULAR BONE; INBRED MICE; IN-VIVO; DIFFERENTIATION; EXPANSION; RECEPTORS; MIGRATION; GROWTH;
D O I
10.1038/nprot.2009.238
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Unlike humans, mouse bone marrow-derived mesenchymal stem cells (MSCs) cannot be easily harvested by adherence to plastic owing to the contamination of cultures by hematopoietic cells. The design of the protocol described here is based on the phenomenon that compact bones abound in MSCs and hematopoietic cells exist in the marrow cavities and the inner interfaces of the bones. The procedure includes flushing bone marrow out of the long bones, digesting the bone chips with collagenase type II, deprivation of the released cells and culturing the digested bone fragments, out of which fibroblast-like cells migrate and grow in the defined medium. The entire technique requires 5 d before the adherent cells are readily passaged. Further identification assays confirm that these cells are MSCs. We provide an easy and reproducible method to harvest mouse MSCs that does not require depletion of hematopoietic cells by sorting or immunomagnetic techniques.
引用
收藏
页码:550 / 560
页数:11
相关论文
共 61 条
[61]   Tumor Necrosis Factor-α Alters the Modulatory Effects of Mesenchymal Stem Cells on Osteoclast Formation and Function [J].
Zhu, Heng ;
Jiang, Xiao-Xia ;
Guo, Zi-Kuan ;
Li, Hong ;
Su, Yong-Feng ;
Yao, Hui-Yu ;
Wang, Xiao-Yan ;
Li, Xiu-Sen ;
Wu, Ying ;
Liu, Yuan-Lin ;
Zhang, Yi ;
Mao, Ning .
STEM CELLS AND DEVELOPMENT, 2009, 18 (10) :1473-1484