The Evaluation of Myelination and the Expression of Neurotrophic Factors in Human Adipose-Derived Stem Cells Inducedinto Schwann- Like Cells at the Presence of Laminin

Background and Aim: Cell-based therapy for repairment of nerve injuryis one of the important strategies in nerve regeneration. Schwann cells(SCs)play a critical role in nerve regeneration, furthermore, SCs secretesneurotrophic factors (NTFs), a main group of polypeptides contributingto the maintenance of neuronal cells and promoting development.However, there is a limitation in obtaining SCs to use. Human Adipose-Derived Stem Cells(hADSCs) are multipotent stem cells and are suitableto use. Laminin is an essential protein in ECM synthesize by SCs. Also.The objective of this study was to induced hADSCs into SC-like cellsand assess the effect of laminin-coated culture on the efficiency of SClikecells differentiation, the myelination potential and the expression of neurotrophic factors.Methods: The human ADSCs were isolated from the adipose tissue, thesecells were plated in pre-induction medium, after neurospheres formation,these cells were re-plated with complete differentiation medium inthe presence and absence of laminin-coated flasks. Then, Schwanncell markers S100β, GFAP and myelin basic proteins (MBP) of SC-likecells were assessed through immunocytochemistry. The quantify genesexpression for Schwann cell markers, MBP and neurotrophic factorslike brain-derived neurotrophic factor (BDNF), ciliary neurotrophicfactor (CNTF), glial cell-derived neurotrophic factor (GDNF) and nervegrowth factor (NGF) was analyzed by the real-time reverse transcriptionpolymerase chain reaction (RT-PCR) technique.Results: The results of this study indicated that the phenotype of thedifferentiated ADSCs was converted to bi-polar or spindle shape inthe absence and presence of laminin. Immunocytochemistry analysisrevealed that the mean percentage of co-markers GFAP/S100β and S100β/MBP positive cells can increase the differentiated rate and myelinogenicpotential of SC-like cells, coating surface by laminin as compared tocontrol group. The genes expression levels of SCs markers, MBP andneurotrophic factors in SC-like cells in two condition was performedby RT-PCR. The findings data demonstrated that gene expression of SCsmarkers and MBP, in addition, neurotrophic factors BDNF and GDNFincrease significantly in the presence of laminin compared to a plasticsurface (P < 0.01). In versus, the expression of NGF was downregulatedsignificantly on laminin-coated plates as compared to that of the absenceof laminin (P < 0.05). Also, the expression of CNTF was upregulated butnot significant.Conclusion: The findings of this present study suggest that the expressionof myelination-related gene and neurotrophic factors genes becomesupregulated in the presence of laminin. Therefore, laminin may providea more appropriate microenvironment for peripheral nerve regenerationand neurodegenerative diseases.

کلیدواژه ها:

Adipose-derived stem cells ، Laminin ، Myelin ، Neurotrophic factors