Journal
HAEMATOLOGICA-THE HEMATOLOGY JOURNAL
Volume 95, Issue 9, Pages 1594-1598Publisher
FERRATA STORTI FOUNDATION
DOI: 10.3324/haematol.2009.019828
Keywords
erythroid; erythropoiesis; CD34
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Funding
- NHS Blood and Transplant (NHSBT)
- BBSRC DTA NHSBT Case Studentship
- NHSBT Wellcome Trust
- National Institute for Health Research [RP-PG-0310-1004] Funding Source: researchfish
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The study of human erythropoiesis in health and disease requires a robust culture system that consistently and reliably generates large numbers of immature erythroblasts that can be induced to differentiate synchronously. We describe a culture method modified from Leberbauer et al. (2005) and obtain a homogenous population of erythroblasts from peripheral blood mononuclear cells (PBMC) without prior purification of CD34(+) cells. This pure population of immature erythroblasts can be expanded to obtain 4x10(8) erythroblasts from lx10(8) PBMC after 13-14 days in culture. Upon synchronized differentiation, high levels of enucleation (80-90%) and low levels of cell death (<10%) are achieved. We compared the yield of erythroblasts obtained from PBMC, CD34(+) cells or PBMC depleted of CD34(+) cells and show that CD34(-) cells represent the most significant early erythroid progenitor population. This culture system may be particularly useful for investigating the pathophysiology of anemic patients where only small blood volumes are available.
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