A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone.
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.
Zhu
Heng
H
Department of Cell Biology, Institute of Basic Medical Sciences, Beijing, China.
Guo
Zi-Kuan
ZK
Jiang
Xiao-Xia
XX
Li
Hong
H
Wang
Xiao-Yan
XY
Yao
Hui-Yu
HY
Zhang
Yi
Y
Mao
Ning
N
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases; D005260:Female; D006412:Hematopoietic Stem Cells / Q000166:cytology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D039902:Multipotent Stem Cells / Q000166:cytology
D017364:Collagenases