[Experimental study of osteogenic induction of fetal mouse liver mesenchymal stem cells in vitro and their biologic attachment properties to true bone ceramic].
OBJECTIVE
To study the culture and purification of the fetal mouse liver mesenchymal stem cells (MSCs) in vitro and to investigate their differentiation potential and the composite ability with true bone ceramic(TBC).


METHODS
The single cell suspension of MSCs was primarily cultured and passaged, which was prepared from the fetal mouse liver; the flow cytometry was applied to detect CD29, CD34, CD44 and CD45. The osteogenic differentiation was induced in chemical inducing system; the osteogenic induction potency was tested. The purified fetal mouse liver MSCs were compounded with TBC covered with collagen type I in vitro and the cell attachment and proliferation to the TBC were observed.


RESULTS
The primary MSCs of fetal mouse liver were easy to culture in vitro. They proliferated well and were easy to subcultured. The proliferation ability of primary and passaged MSCs was similar. Flow cytometric analysis showed the positive results for CD29, CD44 and the negative results for CD34, CD45. After 7 days of induction, the MSCs expressed collagen type I and alkaline phosphatase(ALP) highly. After 14 days of induction, the fixed quantity of ALP increased significantly. After 28 days of induction, calcium accumulation was observed by Von Kossa's staining. Many liver MSCs attached to the surface of TBC.


CONCLUSIONS
The MSCs of the fetal mouse liver can be obtained, subcultured and purified easily. After culturing in chemical inducing system, the MSCs of fetal mouse liver can be successfully induced to osteoblast-like cells, attach to the surface of TBC and proliferate well.
Wang
Jianlong
J
Department of Orthopaedics, the Third Xiangya Hospital, Central South University, Changsha Hunan 410013, P R China. wangjl1972@sina.com
Zhou
Jiangnan
J
Zhao
Ziping
Z
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D018786:Bone Substitutes / Q000378:metabolism; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D002516:Ceramics / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D018960:Hyaluronan Receptors / Q000032:analysis; D019012:Integrin beta1 / Q000032:analysis; D017493:Leukocyte Common Antigens / Q000032:analysis; D008099:Liver / Q000166:cytology* / Q000196:embryology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D010012:Osteogenesis; D011247:Pregnancy; D054457:Tissue Scaffolds
D018952:Antigens, CD34; D018786:Bone Substitutes; D018960:Hyaluronan Receptors; D019012:Integrin beta1; D017493:Leukocyte Common Antigens