Isolation and in vitro expansion of rabbit bone marrow mesenchymal stem cells

Objective This study is designed to explore the isolation,cultivation of bone marrow mesenchymal stem cells(MSCs) in vitro. Methods Rabbit bone marrow was aspirated from the iliac crest sterilely with heparin. Bone marrow mononuclear cells were isolated by gradient density centrifugation with lymphocyte separation medium.Mesenchymal stem cells(MSCs) were then obtained and cultured in completely chemically defined medium.Common cell culture technique and normal methods were used to study the effects on the growth,proliferation,morphology of MSCs in different pre-adsorbed protein coated plastic plate,adherent time and cell density. Results The optimal condition in vitro for growth was pre-adsorbed protein coated on the plastic bottom ,48～72 hours adherent time,(3～9) 10 4 /ml cell density.Spindle cells had a potential ability of proliferation and pluripotent differentiation. Conclusion The optimal condition for isolation and culture of bone marrow MSCs were successfully established.It is a base for further investigation and use of MSCs.