Summary
Recent developments have focused on long-term bone marrow cultures and serum-free media to establish clonal hematopoiesis in vitro. Prior investigations have suggested an important role for such ingredients as transferrin, albumin, lipid, selenite, and potassium provided by serum. These factors, when present, can support clonal growth even in the absence of serum. Results reported here further define the importance of the cations K+ and Li+ in the regulation of in vitro erythropoiesis in the presence of both normal and dialyzed fetal calf serum and ouabain. While K+ proved essential for optimal erythroid colony formation, the presence of Li+ in such cultures reduced optimal colony formation of erythroid stem cells. This evidence suggests that in long-term marrow cultures or serum-free media K+ is essential for successful maintenance of self-renewal of erythroid stem cells and their differentiation, and that the monitoring of K+ levels may prove useful in such cultures. Since in vitro erythropoiesis was reduced in the presence of Li+, the mechanisms controlling differentiation of hematopoietic stem cell may be interpreted to be subject to cation influence.
Similar content being viewed by others
References
Bernstein A, Hunt DM, Crichley V, Mak TM (1976) Induction by ouabain of hemoglobin synthesis in cultured Friend cells. Cell 9:375–381
Chan HSL, Saunders EF, Freedman MH (1980) Modulation of human hematopoiesis by prostaglandins and lithium. J Lab Clin Med 95:125–129
Dexter TM, Testa NG (1976) Differentiation and proliferation of hematopoietic stem cells in culture. In: Prescott DM (ed) Methods in cell biology, vol 14. Academic Press, New York, pp 387–405
Eagle H (1955) Nutrition needs of mammalian cells in tissue culture. Science 122:501–504
Eagle H (1955a) Amino acid metabolism in mammalian cell cultures. Science 130:432–437
Gallicchio VS, Chen MG (1980) Modulation of murine pluripotential stem cell proliferation by lithium carbonate. Blood 56:1150–1152
Gallicchio VS, Chen MG (1981) Lithium influences the proliferation of hematopoietic stem cells. Exp Hematol 9:804–811
Gallicchio VS, Murphy MJ Jr (1979a) In vitro erythropoiesis. II. cytochemical enumeration of erythroid progenitor stem cells (CFU-e and BFU-e) from normal mouse and human hematopoietic tissues. Exp Hematol 7:219–224
Gallicchio VS, Murphy MJ Jr (1979) In vitro erythropoiesis. III. The role of potassium ions in erythroid colony formation. Exp Hematol 7:225–230
Gallicchio VS, Murphy MJ Jr (1981) Erythropoiesis in vitro. IV. Effects of ouabain on erythroid stem cells (CFU-e and BFU-e). Stem Cells 1:30–37
Gregory CJ, McCulloch EA, Till JE (1973) Erythropoietic progenitors capable of colony formation in culture. State of differentiation. J Cell Physiol 81:411–416
Guilbert LG, Iscove NN (1976) Partial replacement of serum by selenite, transferrin, albumin and lecithin in hematopoietic cell cultures. Nature 263:594–595
Gupta RC, Robinson WA, Kurnick JE (1976) Felty's syndrome. Effect of lithium on granulopoiesis. Am J Med 61:26–32
Harker WG, Rothstein F, Clarksen D, Athens JW, MacFarlane JL (1977) Enhancement of colony-stimulating activity production by lithium. Blood 49:263–267
Iscove NN, Melchers F (1978) Complete replacement of serum by albumin, transferrin and soybean lipid in cultures of lipopolysaccharide reactive B lymphocytes. J Exp Med 147:923–933
Levitt LG, Quesenberry PJ (1980) Effect of lithium on murine hematopoiesis in a liquid culture system. N Eng J Med 302:713–719
Mager DL, MacDonald ME, Bernstein A (1979) Growth in high K+ medium induces Friend cell differentiation. Dev Biol 70:268–273
Matoth Y, Ben-Porath (Arkin) E (1959) Effect of erythropoietin on the mitotic rate of erythroblasts in bone marrow cultures. J Lab Clin Med 54:722–725
Quastel MR, Kaplan JG (1970) Lymphocyte stimulation. The effects of ouabain on nucleic acid and protein synthesis. Exp Cell Res 62:407–420
Rothstein G, Clarkson DL, Larsen W, Grosser BI, Athens JW (1978) Effect of lithium on neutrophil mass and production. N Eng J Med 198:178–180
Spitzer G, Verma DS, Barlogie B, Beran MA, Dicke KA (1979) Possible mechanisms of action of lithium on augmentation of in vitro spontaneous myeloid colony formation. Cancer Res 39:3215–3219
Spivak JL, Misiti J, Stuart R, Sharkis SJ, Sensenbrenner LL (1980) Selective suppression and potentiation of hematopoietic precursor cell proliferation by ouabain. Blood 56:315–317
Temin HW, Pierson RW, Dulak NC (1972) In: Rothblat GH, Cristialo J (eds) Growth nutrition and metabolism of cells in culture. Academic Press, New York, pp 47–57
Tepperman AD, Curtis JE, McCulloch EA (1974) Erythropoietin colonies in cultures of human marrow. Blood 44:459–463
Author information
Authors and Affiliations
Additional information
Supported by grants AM-19741, AM-07266 and CA-06519 from the N.I.H. and the Bob Hipple Memorial Committee for Cancer Research
Rights and permissions
About this article
Cite this article
Gallicchio, V.S., Murphy, M.J. Cation influences on in vitro growth of erythroid stem cells (CFU-e and BFU-e). Cell Tissue Res. 233, 175–181 (1983). https://doi.org/10.1007/BF00222241
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00222241