Differentiation Of Human Bone Marrow Mesenchymal Stem Cells Into Motor Neurons In Vitro

Objective To explore the differentiation potentiality of human bone marrow mesenchymal stem cells (hMSCs) into motor neurons in vitro.Methods The hMSCs were isolated from fresh bone marrow by a method of density gradient centrifugation , then amplified and cultivated in a total medium of DMEM/ F12 containing 10 % FBS. hMSCs were divided into two groups in the first passage. In the induction group , hMSCs were pretreated with a total mediem consisting of 10μg/L basic Fibroblast growth factor (bFGF) for 24 h ,and then induced with 0.5μg/ ml ratinoic acid (RA) and 200 ng/ mL sonic hedgehog (Shh) for 10 days. On the other hand , no inductive agents were added in the control group . After the induction , hMSCs were observed using phase-contrast microscopy. Nestin and NSE,the special marks of neural stem cells and neuron cells were detected by immunocytochemistry and the percentages of them were calculated. HB9, the tag of motor neurons were detected by flowcytometry and analyzed by software.Results Under induction conditions, some hMSCs resumed typical neuronal morphology , and the percentages of Nestin,NSE,HB9 positive cells were (25.00±4.47) %,(51.52±4.40)%,(50.04±5.25) % respectively. But in the control group , hMSCs remained unchange morphologically and none of them expressed Nestin,NSE, and the expression of HB9 is less than 5%. There was a significant difference between the two groups(p<0.05) .Conclusions hMSCs can be induced to differentiate into motor neurons in vitro.