In Vitro Hematopoietic Differentiation of Human Embryonic Stem Cells Induced by Co-Culture With Human Bone Marrow Stromal Cells and Low Dose Cytokines

Cell Biol Int. 2005 Aug;29(8):654-61. doi: 10.1016/j.cellbi.2005.03.019.

Abstract

Human embryonic stem (hES) cells randomly differentiate into multiple cell types during embryoid body (EB) development and limited studies have focused on directed hematopoietic differentiation. Here, we report that the treatment of hES cells during EBs development with a combination of low dose hematopoietic cytokines, including stem cell factor (SCF), Flt-3 ligand, vascular endothelial growth factor (VEGF) and human bone marrow stromal cells (hBMSCs), generated cell clusters that contained 8.81% KDR-positive hemangioblasts, 9.94% CD34-positive hematopoietic stem cells and 25.7% CD45-positive mature hematopoietic cells, and expressed hematopoietic genes such as KDR, stem cell leukemia (scl) and runt-related transcription factor 1 (Runx1). We provide the first evidence for the role of the cytokine-hBMSCs combination in promoting hematopoietic differentiation of hES cells, and thus provide the potential for generation of hematopoietic cells, as well as for understanding early developmental events that govern the initiation of hematopoiesis in humans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / metabolism
  • Basic Helix-Loop-Helix Transcription Factors
  • Biomarkers / metabolism
  • Bone Marrow Cells / physiology*
  • Cell Differentiation / drug effects*
  • Cell Lineage
  • Coculture Techniques
  • Colony-Forming Units Assay
  • Core Binding Factor Alpha 2 Subunit
  • Cytokines / pharmacology*
  • DNA-Binding Proteins / metabolism
  • Dose-Response Relationship, Drug
  • Embryo, Mammalian / cytology*
  • Fetus
  • Flow Cytometry
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Proto-Oncogene Proteins / metabolism
  • Stem Cell Factor / metabolism
  • Stem Cells / cytology*
  • Stromal Cells / physiology*
  • T-Cell Acute Lymphocytic Leukemia Protein 1
  • Transcription Factors / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Antigens, CD
  • Basic Helix-Loop-Helix Transcription Factors
  • Biomarkers
  • Core Binding Factor Alpha 2 Subunit
  • Cytokines
  • DNA-Binding Proteins
  • Proto-Oncogene Proteins
  • RUNX1 protein, human
  • Stem Cell Factor
  • T-Cell Acute Lymphocytic Leukemia Protein 1
  • Transcription Factors
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • TAL1 protein, human