NIH Research Festival
Recent advances in the field of cell transformation provide unique opportunities for in vitro human disorder modeling, such as using iPSC-derived neurons for the study of neurological disorders. Although mixed cultures of autologous inflammatory cells and iPS-derived neurons are the best for in vitro modeling of neuroinflammation, it is still challenging due to limited blood sample size. Based on our published method of directly generating neural stem cells from human peripheral CD34 cells, we aimed to develop a convenient method to separate inflammatory cells and generate iPSC and neural stem cells from 10 ml of peripheral blood samples for later constituting autologous neuroinflammation models. We optimized conditions of PBMC separation and CD34 cells purification, and further generated induced neural stem cells (iNSC) and induced pluripotent stem cells (iPSC) simultaneously from CD34 cells transfected with Sendai virus vectors containing Yamanaka factors with the help of selective media. The neural stem cells could be derived within two weeks after transfection and iPSC appeared in three weeks, as confirmed by immunostaining for cell specific markers. Meanwhile, we could still collect PBMCs from the same blood sample for co-culturing with the derived neural cells, thus creating an autologous model of neuroinflammation from as low as 10 ml of human blood.This method has been used in our unit to generate iNSC/iPSC for a variety of neurological disorders. We expect this model can help researchers generate autologous models for neuroinflammatory disorders.
Scientific Focus Area: Stem Cell Biology
This page was last updated on Friday, March 26, 2021