Article

Large-scale production of embryonic red blood cells from human embryonic stem cells.

Einstein Center for Human Embryonic Stem Cell Research, Department of Medicine, Hematology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Experimental Hematology (impact factor: 2.9). 01/2007; 34(12):1635-42. DOI:10.1016/j.exphem.2006.07.003 pp.1635-42
Source: PubMed

ABSTRACT To develop a method to produce in culture large number of erythroid cells from human embryonic stem cells.
Human H1 embryonic stem cells were differentiated into hematopoietic cells by coculture with a human fetal liver cell line, and the resulting CD34-positive cells were expanded in vitro in liquid culture using a three-step method. The erythroid cells produced were then analyzed by light microscopy and flow cytometry. Globin expression was characterized by quantitative reverse-transcriptase polymerase chain reaction and by high-performance liquid chromatography.
CD34-positive cells produced from human embryonic stem cells could be efficiently differentiated into erythroid cells in liquid culture leading to a more than 5000-fold increase in cell number. The erythroid cells produced are similar to primitive erythroid cells present in the yolk sac of early human embryos and did not enucleate. They are fully hemoglobinized and express a mixture of embryonic and fetal globins but no beta-globin.
We have developed an experimental protocol to produce large numbers of primitive erythroid cells starting from undifferentiated human embryonic stem cells. As the earliest human erythroid cells, the nucleated primitive erythroblasts, are not very well characterized because experimental material at this stage of development is very difficult to obtain, this system should prove useful to answer a number of experimental questions regarding the biology of these cells. In addition, production of mature red blood cells from human embryonic stem cells is of great potential practical importance because it could eventually become an alternate source of cell for transfusion.

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Keywords

CD34-positive cells
 
cell number
 
culture large number
 
earliest human erythroid cells
 
erythroid cells
 
fetal globins
 
flow cytometry
 
Globin expression
 
hematopoietic cells
 
large numbers
 
light microscopy
 
liquid culture
 
mature red blood cells
 
nucleated primitive erythroblasts
 
primitive erythroid cells
 
primitive erythroid cells present
 
resulting CD34-positive cells
 
three-step method
 
undifferentiated human embryonic
 
yolk sac