Enrichment of FGF8-expressing cells from neurally induced human pluripotent stem cell cultures

Stem Cell Reports. 2023 Nov 14;18(11):2240-2253. doi: 10.1016/j.stemcr.2023.10.007. Epub 2023 Nov 2.

Abstract

In early vertebrate development, organizer regions-groups of cells that signal to and thereby influence neighboring cells by secreted morphogens-play pivotal roles in the establishment and maintenance of cell identities within defined tissue territories. The midbrain-hindbrain organizer drives regionalization of neural tissue into midbrain and hindbrain territories with fibroblast growth factor 8 (FGF8) acting as a key morphogen. This organizer has been extensively studied in chicken, mouse, and zebrafish. Here, we demonstrate the enrichment of FGF8-expressing cells from human pluripotent stem cells (hPSCs), cultured as attached embryoid bodies using antibodies that recognize "Similar Expression to Fgf" (SEF) and Frizzled proteins. The arrangement of cells in embryoid body subsets of these cultures and the gene expression profile of the FGF8-expressing population show certain similarities to the midbrain-hindbrain organizer in animal models. In the embryonic chick brain, the enriched cell population induces formation of midbrain structures, consistent with FGF8-organizing capability.

Keywords: ESC; FGF8; IL17RD; SEF; iPSC; isthmic organizer; midbrain-hindbrain boundary; organizer; pluripotent stem cell.

MeSH terms

  • Animals
  • Body Patterning
  • Chickens / metabolism
  • Fibroblast Growth Factor 8 / genetics
  • Fibroblast Growth Factors / metabolism
  • Gene Expression Regulation, Developmental
  • Homeodomain Proteins* / metabolism
  • Humans
  • Mesencephalon / metabolism
  • Mice
  • Pluripotent Stem Cells* / metabolism
  • Zebrafish / metabolism

Substances

  • Homeodomain Proteins
  • Fibroblast Growth Factor 8
  • Fibroblast Growth Factors
  • FGF8 protein, human