Large-scale production of embryonic red blood cells from human embryonic stem cells.
OBJECTIVE
To develop a method to produce in culture large number of erythroid cells from human embryonic stem cells.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Olivier
Emmanuel N
EN
Einstein Center for Human Embryonic Stem Cell Research, Department of Medicine, Hematology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Qiu
Caihong
C
Velho
Michelle
M
Hirsch
Rhoda Elison
RE
Bouhassira
Eric E
EE
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D004912:Erythrocytes / Q000166:cytology* / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology / Q000502:physiology; D020869:Gene Expression Profiling; D005914:Globins / Q000235:genetics; D006801:Humans; D008099:Liver / Q000166:cytology / Q000502:physiology; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology
D018952:Antigens, CD34; D005914:Globins