Neural induction of mesenchymal stem cells via small molecule cocktail treatment

Abstract

Mesenchymal Stem Cells (MSCs) are capable of transdifferentiation into cells of the neuronal lineage when cultured in spent media obtained from cultures of neuroblastoma cell lines or when treated with a cocktail of small molecules. In this study, the principal aim was to transdifferentiate MSCs via spent medium from cultures of the neuroblastoma cell line SH-SY5Y and small molecule cocktails, and to determine the neuronal stage achieved upon treatment. To fulfil these aims, MSCs were extracted from umbilical cord and cultured aseptically. MSCs were first transdifferentiated by spent medium or a small molecule cocktail and then further differentiated using different small molecule cocktails in two distinct stages. RNA was extracted from differentiating cells and used to check CD marker (CD34, CD45, CD73, CD90, and CD105), and neuronal marker (Nanog, SOX2, Nestin, Tubb3, NeuN, and TH) expression. Additionally, protein lysates were used for Western blotting for SOX2, NeuN, Actin, Tubulin, HSP27 and HSP90. The MSCs were successfully differentiated towards mature neuronal cells, as indicated by (1) a stark change in cellular morphology whereby the resultant cells greatly resembled neurons, and (2) a significant change in early and late neuronal marker expression as the cells progressed through the induction protocol. Thus, it was concluded that the devised neural induction protocol works in producing neurons. However, further experiments are required to determine which neuronal stage these MSCs achieve upon differentiation with this protocol. This study lays the groundwork for generation of a robust population of mature and functional neuronal cells and has many possible applications in regenerative therapy and research.