Published in Scientific Works. C Series. Veterinary Medicine, Vol. LVIII ISSUE 3
Written by Ilea Ioana Cristina, Pall Emoke, Ciupe Simona, Cenariu M., I.S.Groza
Mesenchymal stem cells (MSCs) are defined as bone marrow derived cells which have the capacity to differentiate into the three classical mesodermal lineages (adypocites, osteoblasts and chondrocytes). Different studies revealed the ability of bone marrow derived MSCs to differentiate into ectodermic lineages including neurons. The aim of our study was to evaluate the multipotency of mouse bone marrow derived MSCs by phenotypic characterization during neuronal induction. Mouse MSCs were isolated from bone marrow by flushing the femurs with α MEM (Gibco) medium supplemented with 1% penicillin-streptomycin (Gibco). Isolated cells were cultured in a propagation medium containing DMEM-F12 medium supplemented with 20% FCS (Gibco), 1% penicillin-streptomycin (Gibco), 5% horse serum (Sigma) and 10μg/5ml MycoZap (Mycoplasma Elimination Reagent, Lonza). For neural induction, cells were cultured in Neurobasal medium supplemented with 0.1mM β-mercaptoethanol and 1% glutamax for 2 weeks. For phenotypic characterization, were evaluated the expression of S-100 protein and neuron specific enolase (NSE) during differentiation. Our results confirmed the multipotency of isolated cells by neuronal differentiation. At 3 days after neurogenic induction, cells morphology changed, appearing star-shaped cells and at day 4 were present specific neuritic networks. At 2 weeks after induction, the immunostaining showed the presence of S-100+ cells, confirming the glial differentiation, as well as NSE+ cells, an indicator of neuronal differentiation.
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