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A Study on Transdifferentiation of Bone Marrow Stromal Cells into Neuronal and Glial-Like Cells In Vitro by Different Inducers

Introduction: There are some evidences to sugges t that bone marrow stromal cells (BMSCs) not only differentiate into mesodermal ce lls, but also adopt the fate of endodermal and ectodermal cell types. BMSCs can be a valuable cell source as an autograft for clinical application involving regeneration of the central nervous system. Bone marrow stromal cell s can be expanded rapidly in vitro and can be differentiated into neuronal- and glial-like cells. In this study, we attempted to devi se a protocol or protocols for the induction of BMSCs into neuroepithelial- and neuroglial-like ce lls . Materials and Methods: Bone marrow was extracted from the femur and tibia of adult rat, and then bone marrow stromal cells with 4 passages were proliferated and cultured and then were ev aluated with fibron ectin by immunocytochemi stry and Oct-a by semi quantitative RT-PCR techniques. Also in this stage expression of Nestin , NF68, GFAP and 04 antibodies respectively markers of neuroepithelial , neuron, astrocytes and oligodendrocytes cell s, were assessed. Rat BMSCs were differentiated by two consequent inductors into neu roepithelial , neuronal and glial-like cells. At pre-induction stage dimethyl sulfoxide (DMSO), p-mercaptoethanol (P:ME) or biotylated hydroxyani sol (BHA) were separately and without fetal bovine serum (FBS) added to alph a minimal essential med ium (o.-:MEM), and then at induction stage the medium was replaced by retinoic acid (RA) and 15% FBS in a-MEM. Four days later, expressions of neuronal and glial markers were assessed. In addition, expression of NeuroD and Oct-4 mRNA were assessed in these cell s. Results: More than 92% of BMSCs was fibronectin posit ive at passage 4. A few percent of BMSCs differentiated into neuroepithelial and neuron-like cells but no astrocyte and oligodendrocyte- like cell s were detected. Oct-4 mRNA was highly expressed in these cell s whil e NeuroD mRNA ex press ion was not detcted. Induction of BMSCs by DMSO-RA differentiated BMSCs into neuroepithe lial and neuronal-like ce lls sig nificantly compare to P:ME-RA and BHA-RA . Transdifferentiation of the treated BMSCs into astrocytes and oligodendrocyte-like cell s was less than 5 %. Induction of BMSCs by DMSO-RA resulted in expression of NeuroD mRNA but Oct -4 mRNA was not expressed in none of treatment groups. Conclusion: Induction of BMSCs by different inducers specially DMSO-RA could highly tran sdifferentiate BMSCs into neuroepithelial and neuronal-like cell s, whereas glial -like cell s transdifferentiation was ve ry low.