间充质干细胞
骨髓
细胞生物学
胶原酶
造血
干细胞
生物
成纤维细胞
干细胞移植修复关节软骨
组织工程
免疫学
细胞培养
化学
成体干细胞
体外
内皮干细胞
生物化学
遗传学
酶
作者
Heng Zhu,Zi‐Kuan Guo,Xiaoxia Jiang,Hong Li,Xiaoyan Wang,Huiyu Yao,Yi Zhang,Ning Mao
出处
期刊:Nature Protocols
[Springer Nature]
日期:2010-02-25
卷期号:5 (3): 550-560
被引量:470
标识
DOI:10.1038/nprot.2009.238
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.
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