Alternative Routes to Induced Pluripotent Stem Cells Revealed by Reprogramming of the Neural Lineage

• NANOG expression can be obtained without E-cadherin in cells of the neural lineage
• Transgene independence and late markers only occur when colonies are E-cadherin+
• NANOG+E-cadherin+ colonies also upregulate cell cycle-related genes
• DOT1L inhibition increases NANOG+E-cadherin+ colony numbers

SummaryDuring the reprogramming of mouse embryonic fibroblasts (MEFs) to induced pluripotent stem cells, the activation of pluripotency genes such as NANOG occurs after the mesenchymal to epithelial transition. Here we report that both adult stem cells (neural stem cells) and differentiated cells (astrocytes) of the neural lineage can activate NANOG in the absence of cadherin expression during reprogramming. Gene expression analysis revealed that only the NANOG+E-cadherin+ populations expressed stabilization markers, had upregulated several cell cycle genes; and were transgene independent. Inhibition of DOT1L activity enhanced both the numbers of NANOG+ and NANOG+E-cadherin+ colonies in neural stem cells. Expressing SOX2 in MEFs prior to reprogramming did not alter the ratio of NANOG colonies that express E-cadherin. Taken together these results provide a unique pathway for reprogramming taken by cells of the neural lineage.