De Novo Generation of Somatic Stem Cells by YAP/TAZ

J Vis Exp. 2018 May 7:(135):57462. doi: 10.3791/57462.

Abstract

Here we present protocols to isolate primary differentiated cells and turn them into stem/progenitor cells (SCs) of the same lineage by transient expression of the transcription factor YAP. With this method, luminal differentiated (LD) cells of the mouse mammary gland are converted into cells that exhibit molecular and functional properties of mammary SCs. YAP also turns fully differentiated pancreatic exocrine cells into pancreatic duct-like progenitors. Similarly, to endogenous, natural SCs, YAP-induced stem-like cells ("ySCs") can be eventually expanded as organoid cultures long term in vitro, without further need of ectopic YAP/TAZ, as ySCs are endowed with a heritable self-renewing SC-like state. The reprogramming procedure presented here offers the possibility to generate and expand in vitro progenitor cells of various tissue sources starting from differentiated cells. The straightforward expansion of somatic cells ex vivo has implications for regenerative medicine, for understanding mechanisms of tumor initiation and, more in general, for cell and developmental biology studies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Acyltransferases
  • Adaptor Proteins, Signal Transducing / genetics*
  • Adaptor Proteins, Signal Transducing / metabolism
  • Adult Stem Cells / cytology*
  • Adult Stem Cells / metabolism
  • Adult Stem Cells / physiology*
  • Animals
  • Cell Cycle Proteins
  • Cell Differentiation / physiology
  • Islets of Langerhans / cytology
  • Mice
  • Pancreas / cytology*
  • Pancreas / metabolism
  • Pancreas / physiology
  • Pancreas, Exocrine / cytology
  • Pancreatic Ducts / cytology
  • Phosphoproteins / genetics*
  • Phosphoproteins / metabolism
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • YAP-Signaling Proteins

Substances

  • Adaptor Proteins, Signal Transducing
  • Cell Cycle Proteins
  • Phosphoproteins
  • Transcription Factors
  • YAP-Signaling Proteins
  • Yap1 protein, mouse
  • Acyltransferases
  • tafazzin protein, mouse