Self-organized formation of developing appendages from murine pluripotent stem cells

Nat Commun. 2019 Aug 23;10(1):3802. doi: 10.1038/s41467-019-11702-y.

Abstract

Limb development starts with the formation of limb buds (LBs), which consist of tissues from two different germ layers; the lateral plate mesoderm-derived mesenchyme and ectoderm-derived surface epithelium. Here, we report means for induction of an LB-like mesenchymal/epithelial complex tissues from murine pluripotent stem cells (PSCs) in vitro. The LB-like tissues selectively differentiate into forelimb- or hindlimb-type mesenchymes, depending on a concentration of retinoic acid. Comparative transcriptome analysis reveals that the LB-like tissues show similar gene expression pattern to that seen in LBs. We also show that manipulating BMP signaling enables us to induce a thickened epithelial structure similar to the apical ectodermal ridge. Finally, we demonstrate that the induced tissues can contribute to endogenous digit tissue after transplantation. This PSC technology offers a first step for creating an artificial limb bud in culture and might open the door to inducing other mesenchymal/epithelial complex tissues from PSCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Morphogenetic Proteins / genetics
  • Bone Morphogenetic Proteins / metabolism
  • Cell Culture Techniques / methods*
  • Ectoderm / cytology
  • Ectoderm / metabolism
  • Embryo, Mammalian
  • Embryonic Development
  • Epithelium / metabolism
  • Female
  • Forelimb / embryology
  • Forelimb / transplantation
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental / physiology
  • Hindlimb / embryology
  • Hindlimb / transplantation
  • Limb Buds / embryology*
  • Limb Buds / transplantation
  • Male
  • Mice
  • Mouse Embryonic Stem Cells / physiology*
  • Mouse Embryonic Stem Cells / transplantation
  • Signal Transduction / physiology
  • Tissue Engineering / methods*

Substances

  • Bone Morphogenetic Proteins