Transdifferentiation Of Adult Rat Bone Marrow Stromal Cells To Neuron-like Cells In Vitro

Objective: To observe the growth and differentiation of the cultured ratbone marrow cells (BMSC). Methods: separate the bone marrow and purify ,culture the bone marrowstromal cells, study the morphology of the cell with phase contrast microscope.Draw the growth curve by MTT method after subculture of the cells. PurifiedBMSC were induced by 2-mercaptoethanol(2-ME)alone or combination ofBDNF with 2-ME. After 6hours incubation, Neuron special endolase (NSE),neurofilament (NF-200) and glial fibrillary acidic protein (GFAP) were detectedby immunohistochemostry ,and the mRNAs of NSE,NF-200,GFAP weredetected with reverse transcriptase-poly merase chain reaction (rt-PCR). Result: Histoimmunochemistry analysis indicated that all the culturedBMSC were CD 44(+), CD54(+) and CD34(-),CD45(-).After induction with2-ME or BDNF alone or Combination of , some BMSC exhibited the phenotypesof neural cells with long axon-like and dendrite-like processes and branches.After 6 hours induction, NSE and NF-200-positive cells increased significantlybut negative in GFAP immunohistochemical staining,the differentiated rates ofBDNF+2-ME, BDNF, 2-ME were 76.5%, 40.5% and 46.0% respectively. Thereare the same in the mRNA, when compared with control groups and other groups3 houers after the treatment. These results suggest that the addition of BDNFand2-ME caused the maximal variation. the difference among three groups weresignificant(p<0.01). Conclusion: A great number of BMSC with high purity could be obtained byculture in vitro. Combination of 2-ME with BDNF may synergistically promotethe transformation of BMSC and differentiation into neuron-like cells .