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Journal of Iranian Anatomical Sciences. 2009; 6 (25-26): 525-536
in Persian | IMEMR | ID: emr-91771

ABSTRACT

There are some evidences to suggest that bone marrow stromal cells [BMSCs] not only differentiate into mesodermal cells, 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 cells can he expanded rapidly in vitro and can he differentiat into neuronal- and glial-like cells. In this study, we attempted to devise a protocol or protocals for the induction of BMSCS into neuroepithelial- and neuroglial-like cells. 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 evaluated with fibronectin by immunocytochemistry and Oct-4 by semi quantitative RT-PCR techniqucs. Also in this stage expression of Nestin. NF68, GFAP and 04 antibodies respectively markers of neuroepithelia1, neuron astrocytes and oligodendrocytes cells, were assessed. Rat BMSCs were differentiatec; by two consequent indductors into neuroepithelial. neuronal and glial-like cells. At pre-induction stage dimethyl sulfoxide [DMSO], beta-mercaptoethanol [[3ME] or biotylated hydroxyanisol [BHA] were separattly and without fetal bovine serum [betaBS] addled to alpha minimal essential medium [alfa-MEM], and then a induction stage medium was replaced by retinoic acid [RA] and 15% FBS in alfa-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 cells. More than 92% of BMSCs was fibronectin positive at passage 4. A few percent of BMSCs differentiated into neuroepithelial and neuron-like cells but no astrocyte and oligoclendiocyte-like cell were detected. Oct-4 mRNA was highly expressed in these cells while NeuroD mRNA expression was not detected Induction of BMSCs by DMSO-RA differentiated BMSCs into neuroepithelial and neuronal-like cell significantly compare to betaME-RA and BHA-RA. Transdifferentiation of the treated BMSCs into astrocytes and oligodendrocyte-like cells was less than 5%. Indluction of BMSCs by DMSO-RA resulted in expression of NeuroD niRNA but Oct-4 mRNA was not expressed in none of treatment groups. Induction of BMSCs by different inducers specially DMSO-RA could highly transdifferentiate BMSCs into neuroepithelial and neuronal-like cells, whereas glial-like cells transdifiŠrentiation was very low


Subject(s)
Animals, Laboratory , Bone Marrow Cells , Stromal Cells , In Vitro Techniques , Neuroepithelial Cells , Neuroglia , Rats
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