Generation of Integration-Free Porcine Induced Neural Stem Cells Using Sendai virus

Abstract

Background The reprogramming of cells to induced neural stem cells (iNSCs), faster and safer to generate than induced pluripotent stem cells, holds tremendous promise for disease modeling and personalized cell-based therapies for neurological diseases. Porcine iNSCs (piNSCs) may serve as a disease model for human medicine, as pigs are one of the most successful large animal models in biomedical research. Thus, this study aimed to establish safe and efficient integration-free piNSC lines.Methods The integration-free piNSC lines were generated by reprogramming porcine fibroblasts using the Sendai virus (SeV).Results Here we report the successful generation of integration-free piNSC lines using the SeV, with a reprogramming efficiency of 0.4%. The piNSCs can be expanded for up to 40 passages and express high levels of NSC markers (PAX6, NESTIN, and SOX2). They can produce neurons and glia, expressing TUJ, MAP2, TH, and GFAP. No induced pluripotent stem cells developed during reprogramming, and the established piNSCs did not express OCT4. Hence, the SeV can reprogram porcine fibroblast without first going through an intermediate pluripotent stage.Conclusions With the SeV approach, we generated integration-free piNSCs that may be used to assess the efficacy and safety of iNSC-based clinical translation in humans.