In Vitro Differentiation Of Human Umbilical Cord Derived Mesenchymalstem Cells Into Neural Stem Cells

Objective: Human umbilical-cord derived mesenchymal stem cells(UC-MSCs)are pluripotent adult stem cells and present in Wharton's jelly within the UC.UC-MSCs are reported have self-renewal capacity,multi-lineage differentiation potential and the valuable functions of immunomodulatory and paracrine.UC-MSCs have been used for treatment of various neurological diseases.Growing data showed that UC-MSCs can transdifferentiate into neurons and glial cells,but so far a quick and efficient induction method of these cells into neural stem/progenitor cells.In this study,UC-MSCs were induced to form neurosphere-like structures under standard neurobasal medium and then we investigated the biological characteristics of neurospheres derived from UCMSCs.Methods: First of all,UC-MSCs were isolated from umbilical cord and cultured in standard medium contained serum.Secondly,UC-MSCs were induced to form neurosphere-like structures under standard culture condition used for neurospheres(DMED/F12,EGF,b FGF,B27 and N2).And then the self-renewal capacity of UC-MSCs-neurospheres were analyzed by secondary neurosphere formation assay.Neurogenic differentiation was assayed by immunofluorescene staining for neural and glial specific protein expression.The expression of genes were analyzed using quantitative real-time PCR.Next,UC-MSCs and UC-MSCs derived neurospheres(24 hours and 72 hours post-induction)were subjected to the multi-color flow cytometry analysis.Finally,cytokines of UC-MSCs before and after neurosphere formation were assayed by Raybio cytokine microarray analysis.Results: In this study,UC-MSCs were induced to form neurosphere-like structures under standard culture condition used for neurospheres(DMED/F12,EGF,b FGF,B27 and N2)within 12-24 hours.These spheres can self-renew to form secondary spheres and can be readily induced to become neurons and glial cells.Realtime PCR indicated upregulated expression of multiple genes important for neural stem cell self-renewal and multipotency.Flow cytometry analysis showed that UC-MSCs cell surface marker CD13,CD44,CD90 and CD105 significantly down-regulated starting from 72 hours after the induction.Conclusion: Our data showd that NSCs can be efficiently produced from UCMSCs in culture and that these cells may provide an ideal cell source for neuroreplacement therapies with various neurological diseases.