Induction of bone marrow stromal cells into cholinergic-like cells by nerve growth factor

ABSTRACT

Bone marrow stromal cells [BMSC] are used as a source for cell therapy in different model for neurological disorder such as stroke and spinal cord injury. However, the transdifferentiation of BMSC into cholinergic phenotype requires more investigation. BMSC were isolated from adult rats, pre-induced with [beta-mercaptoethanol [BME] and followed by nerve growth factor [NGF] induction. Neurofilaments of 68 kDa, 160 kDa and 200 kDa [NF-200, NF-160 and NF-68, respectively] immuno-staining were used for evaluating the transdifferentiation of BMSC into neuronal phenotype. The percentage of neurofilaments immuno-reactive cells was applied in order to evaluate the results at the pre-induction and the induction stages. Also, NeuroD and Oct-4 expressions, using RT-PCR, were used in assessing the progression of BMSC into neuronal lineage. Choline acetyltransferase immuno-reactive cells were used for estimating the percentage of cholinergic neuronal phenotype. Immuno-staining with anti-microtubule-associated protein-2 [MAP-2] and anti-synapsin-I antibodies was done in order to evaluate cell tendency for synaptogenesis. The yield of cholinergic neurons with BME as pre-inducer and NGF as inducer was 80%. Also, NF-200, NF-160, NF-68, MAP-2 and synapsin-I were detected in the transdifferentiated cells. RT-PCR showed the expression of NeuroD, while Oct-4 was not detected. BME as pre-inducer and NGF as inducer for BMSC transdifferentiation into cholinergic phenotype are potential sources in traumatic injury therapy in the central nervous system