Lentivirus generation for improved oligodendrocyte differentiation
Despite the plethora of research on the central nervous system (CNS), we know surprisingly little about oligodendrocytes–the primary cell affected in diseases like multiple sclerosis. This is in part due to the longer time it takes for these cells to mature (over 100 days) compared to other cell types starting from induced pluripotent stem cells (iPSCs). However, recent studies have shown that by transducing iPSCs with three transcription factors (SOX10, OLIG2, and NKX6.2) the duration can be shortened to less than 30 days. We sought to streamline a methodology for oligodendrocyte differentiation for downstream research. We began by using E. coli to grow the plasmid containing our three transcription factors respectively. Once our plasmid was isolated and its genetic composition was confirmed, we packaged it into a lentivirus vehicle using 293T cells. The viral titer was confirmed using qPCR technology. We have begun iPSC transduction and expect the process to be completed in the next 21 days. We hope researchers can take advantage of this methodology to study the impact of disease on oligodendrocytes. For future studies, this methodology could be used to generate oligodendrocytes for high throughput drug testing and to study intercell communication to better inform researchers about significant changes in this abundant cell type.