Study on amplification of bone marrow mesenchymal stem cells with serum from different sources in vitro

Abstract

Objective To investigate whether autologous serum could replace fetal calf serum for the expansion of human human bone marrow mesenchymal stem cells(BMSCs)in vitro and establish a protocol of culturing BMSCs.Methods BMSCs were separated from human bone marrow by modified ficoll density gradient centrifugation combined with adherence culture method and respectively cultured by DMEM/F12 with 10%umbilical cord blood serum,10%fetal calf serum,human AB blood serum,and autologous serum in vitro. The morphology and its growth character of the BMSCs were studied with the phase-contrast microscope.Differentiation potency through condition medium directional induction were observed. The expression of cell surface antigens was detected by flow cytometer,immunocytochemistry. Results There were no significant differences between four groups in morphological changes. BMSCs were induced to differentiate into neuron like cells by growth factor. The induced cells expressed NSE,NF,GFAP after induction. Conclusion High purity and active BMSCs are cultured by autoserum,which grow prosperously and maintain its biological characters for at least 5 passages. Key words: BMSCs; Autologous serum; Differentiation