The effect of hypoxia and stem cell source on haemoglobin switching

Br J Haematol. 2005 Feb;128(4):562-70. doi: 10.1111/j.1365-2141.2004.05336.x.

Abstract

This study investigated whether relative changes that accompany the naturally occurring shifts in haematopoietic sites during human development play a role in haemoglobin (Hb) switching or whether Hb switching is innately programmed into cells. CD34(+)/Lineage(-) haematopoietic stem/progenitor cells (HSCs) were isolated from human fetal liver (F-LVR), cord blood (CB), and adult bone marrow (ABM), and the Hb was characterized by flow cytometry on cultures that generated enucleated red cells. All feeder layers (stroma from F-LVR, ABM, and human fetal aorta) enhanced cell proliferation and erythropoiesis but did not affect Hb type. HSCs from CB and F-LVR generated the same Hb profile under normoxia and hypoxia. HSCs from ABM had single-positive HbA and double-positive HbA and HbF cells at normoxia and almost entirely double-positive cells at hypoxia. Further characterization of these ABM cultures was determined by following mRNA expression for the transcription factors erythroid Kruppel-like factor (EKLF) and fetal Kruppel-like factor (FKLF) as a function of time in cultures under hypoxia and normoxia. The erythroid-specific isoform of 5-amino-levulinate synthase (ALAS2) was also expressed under hypoxic conditions. We conclude that Hb switching is affected by the environment but not all HSCs are preprogrammed to respond.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Cell Culture Techniques / methods
  • Cell Differentiation / physiology
  • Cell Division / physiology
  • Cell Hypoxia / physiology
  • Cell Lineage / physiology
  • Cells, Cultured
  • Culture Media, Serum-Free
  • Erythropoiesis / physiology*
  • Fetal Blood / cytology
  • Fetal Hemoglobin / metabolism*
  • Hematopoietic Stem Cells / cytology*
  • Hemoglobin A / metabolism
  • Humans
  • Liver / cytology
  • Liver / embryology
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Culture Media, Serum-Free
  • Hemoglobin A
  • Fetal Hemoglobin