The majority of the in vitro erythroid expansion potential resides in CD34(-) cells, outweighing the contribution of CD34(+) cells and significantly increasing the erythroblast yield from peripheral blood samples.
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 1x10(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.
van den Akker
Emile
E
Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Clifton, Bristol, BS81TD, United Kingdom.
Satchwell
Timothy J
TJ
Pellegrin
Stephanie
S
Daniels
Geoff
G
Toye
Ashley M
AM
D018952:Antigens, CD34 / Q000032:analysis*; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D004900:Erythroblasts / Q000166:cytology*; D004920:Erythropoiesis; D006801:Humans
D018952:Antigens, CD34