In vitro mass production of human erythroid cells from the blood of normal donors and of thalassemic patients.
We describe a new two-step culture method for mass production in vitro of erythroid cells from either CD34+ (10(5) cells/mL) or light-density (10(6) cells/mL) cells purified from the blood of normal donors and thalassemic patients. The method includes (i) culture of the cells in the presence of dexamethasone and estradiol (10(-6) M each) and (ii) the growth factors SCF (50 ng/mL), IL-3 (1 ng/mL), and EPO (1 U/mL). In their proliferative phase, these cultures generated approximately 1.2 x 10(7) erythroblasts for each milliliter of blood collected from normal donors or thalassemic patients. They were composed mostly (90%) of CD45(low)/glycophorin (GPA)(neg)/CD71(1ow) cells at day 7, 50-60\% of which became CD45(neg)/GPA+/CD71high by days 15-20. However, when cells from days 7 to 12 of the proliferative phase were transferred in differentiation medium containing EPO and insulin, they progressed to mature erythroblasts (g90% benzidine(pos) and CD45(neg)/GPA+/CD71medium) in 4 days. Because of the high number of erythroid cells that are generated from modest volumes of blood, this method will prove useful in donor-specific studies of erythroid differentiation.
Migliaccio
Giovanni
G
Laboratory of Biochimica Clinica, Istituto Superiore Sanità, Rome, Italy. migliagi@iss.it
Di Pietro
Roberta
R
di Giacomo
Viviana
V
Di Baldassarre
Angela
A
Migliaccio
Anna Rita
AR
Maccioni
Liliana
L
Galanello
Renzo
R
Papayannopoulou
Thalia
T
D018952:Antigens, CD34; D001773:Blood Cells / Q000166:cytology* / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002455:Cell Division; D003907:Dexamethasone / Q000494:pharmacology; D015672:Erythroid Precursor Cells / Q000166:cytology* / Q000276:immunology; D004958:Estradiol / Q000494:pharmacology; D005914:Globins / Q000032:analysis; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D013789:Thalassemia / Q000097:blood*
D018952:Antigens, CD34; D006133:Growth Substances; D004958:Estradiol; D003907:Dexamethasone; D005914:Globins