A Novel Serum-Free Monolayer Culture for Orderly Hematopoietic Differentiation of Human Pluripotent Cells via Mesodermal Progenitors

PLoS One. 2011;6(7):e22261. doi: 10.1371/journal.pone.0022261. Epub 2011 Jul 27.

Abstract

Elucidating the in vitro differentiation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells is important for understanding both normal and pathological hematopoietic development in vivo. For this purpose, a robust and simple hematopoietic differentiation system that can faithfully trace in vivo hematopoiesis is necessary. In this study, we established a novel serum-free monolayer culture that can trace the in vivo hematopoietic pathway from ES/iPS cells to functional definitive blood cells via mesodermal progenitors. Stepwise tuning of exogenous cytokine cocktails induced the hematopoietic mesodermal progenitors via primitive streak cells. These progenitors were then differentiated into various cell lineages depending on the hematopoietic cytokines present. Moreover, single cell deposition assay revealed that common bipotential hemoangiogenic progenitors were induced in our culture. Our system provides a new, robust, and simple method for investigating the mechanisms of mesodermal and hematopoietic differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD34 / metabolism
  • Biomarkers / metabolism
  • Blood Cells / cytology
  • Blood Cells / drug effects
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects*
  • Cell Line
  • Cell Lineage / drug effects
  • Clone Cells
  • Culture Media, Serum-Free / pharmacology*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects
  • Embryonic Stem Cells / metabolism
  • Hematopoiesis / drug effects
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Leukocyte Common Antigens / metabolism
  • Mesoderm / cytology*
  • Mesoderm / drug effects
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / metabolism
  • Stromal Cells / cytology
  • Vascular Endothelial Growth Factor Receptor-2 / metabolism

Substances

  • Antigens, CD34
  • Biomarkers
  • Culture Media, Serum-Free
  • Vascular Endothelial Growth Factor Receptor-2
  • Leukocyte Common Antigens