A ROCK inhibitor permits survival of dissociated human embryonic stem cells

Nat Biotechnol. 2007 Jun;25(6):681-6. doi: 10.1038/nbt1310. Epub 2007 May 27.

Abstract

Poor survival of human embryonic stem (hES) cells after cell dissociation is an obstacle to research, hindering manipulations such as subcloning. Here we show that application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency (from approximately 1% to approximately 27%) and facilitates subcloning after gene transfer. Furthermore, dissociated hES cells treated with Y-27632 are protected from apoptosis even in serum-free suspension (SFEB) culture and form floating aggregates. We demonstrate that the protective ability of Y-27632 enables SFEB-cultured hES cells to survive and differentiate into Bf1(+) cortical and basal telencephalic progenitors, as do SFEB-cultured mouse ES cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / administration & dosage*
  • Animals
  • Cell Differentiation / drug effects*
  • Cell Survival / drug effects*
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism*
  • Humans
  • Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Mice
  • Protein-Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein-Serine-Threonine Kinases / metabolism*
  • Pyridines / administration & dosage*
  • rho-Associated Kinases

Substances

  • Amides
  • Intracellular Signaling Peptides and Proteins
  • Pyridines
  • Y 27632
  • Protein-Serine-Threonine Kinases
  • rho-Associated Kinases