Primary culture and biological characteristics of bone marrow mesenchymal stem cells from a rabbit / 中国组织工程研究

ABSTRACT

BACKGROUND:Rabbit bone marrow mesenchymal stem cels as a kind of adult stem cels with strong proliferation and multilineage differentiation potential exhibit a tremendous application potential in tissue engineering and biological therapy. OBJECTIVE:To in vitro culture, proliferate and identify rabbit bone marrow mesenchymal stem cels and to observe cel biological characteristics. MEHTODSBone marrow of rabbits was extracted under sterile conditions to separate bone marrow mesenchymal stem cels using the whole bone marrow adherence method and Percol density gradient centrifugation method. Afterwards, the cels were purified and proliferated using differential adherence method. Morphology and growth pattern of cels were observed under microscope, and expression of cel surface antigen markers was detected by flow cytometry. RESULTS AND CONCLUSION:Rabbit bone marrow mesenchymal stem cels presented with short adherent time and fast growth. After passage and purification, impurities cel counts were decreased. Primary cels presented with triangular, fusiform and spindly shapes. Passage 5 cels with single shape showed the typical polar swirling growth, and could not express CD34 and CD45, but expressed CD29 and CD44. These findings indicate that the cels cultured using the whole bone marrow adherence method and Percol density gradient centrifugation method possess stem cel characteristics in morphology, surface markers and multilineage differentiation, which have been identified as bone marrow mesenchymal stem cels by flow cytometry.