Methylcobalamin induces differentiation of rat bone mesenchymal stem cells into neuron-like cells in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND:Currently, transplantation of bone marrow mesenchymal stem cel s into the spinal cord is very limited to the recovery of animals fol owing spinal cord injury. Methylcobalamin is a common drug for the treatment of neurological diseases and injuries, but its effects on bone marrow mesenchymal stem cel s are unclear. OBJECTIVE:To study the feasibility of bone marrow mesenchymal stem cel s differentiating into neuron-like cel s induced by methylcobalamin in vitro and to observe the cel viability and proliferation of differentiated cel s. Methods:Rat bone marrow mesenchymal stem cel s were isolated, cultured and purified by density gradient centrifugation and adherent culture. The fourth to fifth generation of bone marrow mesenchymal stem cel s were treated for 24, 48 and 72 hours with different concentrations (25, 50 and 100 mg/L) of methylcobalamin. The morphological changes and cel growth were continuously observed under an inverted phase constract microscope. The viability of induced cel s was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The expressions of Nestin and neuron-specific enolase were identified by reverse transcription PCR and western blot. RESULTS AND CONCLUSION:Most of bone marrow mesenchymal stem cel s could differentiate into neuron-like cel s after induction with methylcobalamin. The expressions of Nestin and neuron-specific enolase were up-regulated after 48 hours of methylcobalamin treatment at different concentrations, especial y after treatment with 100 mg/L methylcobalamin. Similarly, the expressions of Nestin and neuron-specific enolase could be increased significantly after 100 mg/L methylcobalamin treatment for 24, 48 and 72 hours, especial y for 72 hours. It is indicated that methylcobalamin can induce bone marrow mesenchymal stem cel s differentiating into neuron-like cel s, and the optimal concentration of methylcobalamin is 100 mg/L.