TY - JOUR
AU - Rachmilewitz, E. A.
AB - Abstract Most studies of erythropoiesis in vitro have employed cloning methods in semisolid medium. We have recently described a two-step liquid culture procedure that supports the proliferation and differentiation of human erythroid progenitors. In the present study, we have modified the procedure to allow large-scale cultures of erythroid cells derived from normal donors. The culture is divided into two phases. In the first phase, which is erythropoietin (Epo) independent, the early erythroid progenitors multiply and differentiate. In the second, Epo-dependent phase, they mature into orthochromatic normoblasts and enucleated erythrocytes. Using this procedure, erythroid cell yield reached 7.5 × 106/ml and a total of 7 × 108 cells could be harvested per blood unit. A comparison of the growth of erythroid cells in liquid culture to their colony growth in semisolid culture indicated that cell growth was superior: 1) in liquid culture in terms of cell yield per originally cultured mononuclear cell, 2) per ml culture and per culture surface area and in the purity of the resultant erythroid cell population. In addition, it permits easier manipulation of the culture condition and components and sampling of > 1 × 107 cells at each maturation stage subsequent to the proerythroblast stage. This liquid culture procedure might provide an important experimental tool for studying erythroid cell development. Erythropoiesis, Two-phase liquid culture, Large-scale culture References 1 Friend C , Scher W, Holland J, Sato J. Hemoglobin synthesis in murine virus induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethylsulfoxide . Proc Natl Acad Sci USA 1971 ; 68 : 378 – 382 . Google Scholar Crossref Search ADS PubMed WorldCat 2 Lozzio CB , Lozzio BB. Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome . Blood 1975 ; 45 : 321 – 334 . Google Scholar Crossref Search ADS PubMed WorldCat 3 Stephenson JR , Axelrad AA, McLeod DL, Shreeve MM. Induction of colonies of hemoglobin synthesizing cells by erythropoietin . Proc Natl Acad Sci USA 1971 ; 68 : 1542 – 1546 . Google Scholar Crossref Search ADS PubMed WorldCat 4 Iscove NN . Erythropoietin-independent stimulation of early erythropoiesis in adult marrow cultures by conditioned media from lectin stimulated mouse spleen cells. In: Golde DW, Cline MJ, Metcalf D, Fox CF, eds. Hemopoietic Cell Differentiation . San Diego : Academic Press , 1978 : 37 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 5 Metcalf M . Hemopoietic colonies: in vitro cloning of normal and leukemic cells . New York : Springer-Verlag , 1977 . Google Scholar Crossref Search ADS Google Preview WorldCat COPAC 6 Fibach E , Manor D, Oppenheim A, Rachmilewitz EA. Proliferation and maturation of human erythroid progenitors in liquid culture . Blood 1989 ; 73 : 100 – 103 . Google Scholar Crossref Search ADS PubMed WorldCat 7 Clarke BJ , Housman D. Characterization of an erythroid precursor cell of high prolifer-ative capacity in normal peripheral blood . Proc Natl Acad Sci USA 1977 ; 74 : 1105 – 1109 . Google Scholar Crossref Search ADS PubMed WorldCat 8 Ogawa M , Owen CG, O'Dell RF, Hara H, MacEachern MD. Circulating erythropoietic precursors assessed in culture: characterization in normal men and patients with hemoglobinopathies . Blood 1977 ; 50 : 1081 – 1092 . Google Scholar Crossref Search ADS PubMed WorldCat 9 Myers CD , Katz FE, Joshi G, Millar JL. A cell line secreting stimulating factors for CFU-GEMM culture . Blood 1984 ; 64 : 152 – 155 . Google Scholar Crossref Search ADS PubMed WorldCat 10 Orkin SH , Harosi FI, Leder P. Differentiation of erythroleukemic cells and their somatic hybrids . Proc Natl Acad Sci USA 1975 ; 72 : 98 – 102 . Google Scholar Crossref Search ADS PubMed WorldCat 11 Eaves CJ , Eaves AC. Erythropoietin (Ep) dose-response curves for three classes of erythroid progenitors in normal human marrow and in patients with polycythemia vera . Blood 1978 ; 52 : 1196 – 1210 . Google Scholar Crossref Search ADS PubMed WorldCat 12 Hale G , Bright S, Chumbley G, et al. Removal of T cells from bone marrow for transplantation: a monoclonal anti-lymphocyte antibody that fixes human complement . Blood 1983 ; 62 : 873 – 882 . Google Scholar Crossref Search ADS PubMed WorldCat 13 Borel JF , Ryffel B. The mechanism of action of cyclosporin; a continuing puzzle. In: Schnidler R, ed. Cyclosporin in autoimmune diseases . Heidelberg : Springler-Verlag , 1985 : 24 – 32 . Google Scholar Crossref Search ADS Google Preview WorldCat COPAC This content is only available as a PDF. © 1991 AlphaMed Press This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)
TI - Growth of human normal erythroid progenitors in liquid culture: A comparison with colony growth in semisolid culture
JO - Stem Cells
DO - 10.1002/stem.5530090108
DA - 1991-01-01
UR - https://www.deepdyve.com/lp/oxford-university-press/growth-of-human-normal-erythroid-progenitors-in-liquid-culture-a-g02QpXmhsT
SP - 57
EP - 64
VL - 9
IS - 1
DP - DeepDyve
ER -