Induction of integration-free human-induced pluripotent stem cells under serum- and feeder-free conditions

In Vitro Cell Dev Biol Anim. 2020 Jan;56(1):85-95. doi: 10.1007/s11626-019-00412-w. Epub 2019 Nov 25.

Abstract

Human-induced pluripotent stem cells (hiPSCs) have shown great potential toward practical and scientific applications. We previously reported the generation of human dental pulp stem cells using non-integrating replication-defective Sendai virus (SeVdp) vector in feeder-free culture with serum-free medium hESF9. This study describes the generation of hiPSCs from peripheral blood mononuclear cells to increase the donor population, while reducing biopsy invasiveness. From 6-d-old primary culture of peripheral blood mononuclear cells (PBMCs) with IL-2, hiPSCs were established using SeVdp(KOSM)302L with recombinant Laminin-511 E8 fragments under serum-free condition. The established PBMC-derived hiPSCs showed pluripotency and differentiation ability both in vivo and in vitro. In addition, we evaluated microarray data from PBMC- and dental pulp-derived hiPSCs. These hiPSCs will be beneficial for characterizing the molecular mechanisms of cellular differentiation and may provide useful substrates for developing cellular therapeutics.

Keywords: Human-induced pluripotent stem cells; Peripheral blood mononuclear cells; Reprogramming efficiency; hESF9 serum-free defined media.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Cell Line
  • Cellular Reprogramming
  • Culture Media, Serum-Free / pharmacology*
  • Feeder Cells / cytology*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Leukocytes, Mononuclear / cytology

Substances

  • Culture Media, Serum-Free
  • Alkaline Phosphatase