Generation of neural progenitor cells from porcine-induced pluripotent stem cells

J Tissue Eng Regen Med. 2020 Dec;14(12):1880-1891. doi: 10.1002/term.3143. Epub 2020 Nov 23.

Abstract

In this study, porcine embryonic fibroblasts (pEFs) were reprogrammed into porcine-induced pluripotent stem cells (piPSCs) using either human or mouse specific sequences for the OCT4, SOX2, c-Myc, and KLF4 transcription factors. In total, three pEFs lines were reprogrammed, cultured for at least 15 passages, and characterized regarding their pluripotency status (alkaline phosphatase expression, embryoid body formation, expression of exogenous and endogenous genes, and immunofluorescence). Two piPSC lines were further differentiated, using chemical inhibitors, into putative neural progenitor-like (NPC-like) cells with subsequent analyses of their morphology and expression of neural markers such as NESTIN and GFAP as well as immunofluorescent labeling of NESTIN, β-TUBULIN III, and VIMENTIN. NPC-like cells were positive for all the neural markers tested. These results evidence of the generation of porcine NPC-like cells after in vitro induction with chemical inhibitors, representing an adequate model for future regenerative and translational medicine research.

Keywords: cellular reprogramming; induced pluripotent stem cells (iPSC); neural progenitor cells; swine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Animals
  • Biomarkers / metabolism
  • Cell Differentiation* / genetics
  • Cell Line
  • Cell Shape
  • Cellular Reprogramming
  • Embryoid Bodies / cytology
  • Gene Expression Regulation
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Kruppel-Like Factor 4
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / metabolism
  • Neurons / cytology
  • Swine

Substances

  • Biomarkers
  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Alkaline Phosphatase