Complete replacement of serum in primary cultures of erythropoietin-dependent red cell precursors (CFU-E) by albumin, transferrin, iron, unsaturated fatty acid, lecithin and cholesterol

Iscove, Norman N.; Guilbert, Larry J.; Weyman, Christine

doi:10.1016/0014-4827(80)90476-0

Cited by 243 publications

(92 citation statements)

References 13 publications

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“…1, left curve). The doseresponse curve of Ep on CFU-e colony formation in adult bone marrow fits very well with the curve in [7] obtained with the same culture system.…”

Section: Resultssupporting

confidence: 73%

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A new candidate for the regulation of erythropoiesis

1982

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The effect of pure human insulin-like growth factor I (IGF I) on the colony formation of late stage erythroid precursor cells (CFU-e) from fetal mouse liver and adult bone marrow was studied in a serumfree culture system. We found that IGF I in physiological concentrations stimulated erythroid colony formation. The combined effect of IGF I and erythropoietin was smaller than the sum of their single effects. The number of colonies induced by IGF I was linearly dependent on the number of plated cells. Our results indicate that IGF I is the first clearly defined mitogen that stimulates the late stages of erythroid differentiation independently of erythropoietin. Insulin-like growth factor Somatomedin Erythroid differentiation Erythroid precursor cellCell structure

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“…1, left curve). The doseresponse curve of Ep on CFU-e colony formation in adult bone marrow fits very well with the curve in [7] obtained with the same culture system.…”

Section: Resultssupporting

confidence: 73%

“…Bone marrow cell suspensions were prepared from femurs of adult female mice. The test procedure was essentially that in [6] except that fetal bovine serum was replaced by a serum-free incubation medium according to [7]. Cells were cultured in 4-well dishes (Greiner/FRG).…”

Section: Erythroid Colony Assaymentioning

confidence: 99%

A new candidate for the regulation of erythropoiesis

1982

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“…7) have contributed to the development of the concept of hormonally defined media and have shown how useful these media can be for analyzing the controls of proliferation and differentiation at the molecular level. Iscove and co-workers (8)(9)(10) have applied this concept to the culture ofmurine hemopoietic cells.…”

mentioning

confidence: 99%

Fibrinogen and its fragment D stimulate proliferation of human hemopoietic cells in vitro.

Hatzfeld¹,

Hatzfeld²,

Maigne³

1982

Proc. Natl. Acad. Sci. U.S.A.

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Purified fibrinogen at concentrations of3-30 nM has been found to stimulate continuous growth ofhuman lymphoid and myeloid cell lines under serum-free conditions. A strong proliferative response resulted from the synergism elicited by the addition of fibrinogen to transferrin-supplemented medium. This effect was observed with the pre-B-cell line Raji, the T lymphomaderived JM, and the monocytic cell line U 937, either at high or low cell densities. With the promyelocytic cell line HL 60, fibrinogen did not shorten the doubling time of cultures seeded at high cell densities (2 x 105 cells per ml). However, at cell densities lower by 2 orders of magnitude and in the same medium, it promoted growth with a doubling time similar to that obtained at high cell concentrations. Fibrinogen also was found to increase the plating efficiency and colony size when human bone marrow cells were cultured in semisolid medium containing serum. In long-term bone marrow liquid cultures without fibrinogen, colony-forming cells were no longer detected after 6 weeks. In those cultured with fibrinogen, -50 granulocyte-macrophage colonies per 105 cells were obtained after 6 weeks, and 10, after 12 weeks. Purified fibrinogen fragment D possessed a stimulating activity similar to that of the intact fibrinogen molecule. This fragment cannot form fibrin, thus eliminating fibrin as a source of the mitogenic effect.The use of serum in tissue culture hinders the study of the regulation of cellular proliferation (1)(2)(3)(4)(5)(6) because the roles and interactions amongst various growth factors are difficult to analyze in nondefined media. Sato and his group (for a review see ref. 7) have contributed to the development of the concept of hormonally defined media and have shown how useful these media can be for analyzing the controls of proliferation and differentiation at the molecular level. Iscove and co-workers (8-10) have applied this concept to the culture ofmurine hemopoietic cells.A minimal medium is composed of defined factors that are all necessary in a particular combination for the growth of a particular cell type; removal ofeach factor individually prevents growth. These media facilitate the study of each growth requirement and its effects. By designing minimal media containing purified plasma components, we have analyzed their possible roles in the control of human hemopoietic cell proliferation (11).Fibrinogen is a major component of plasma, where its concentration is about 3 mg/ml. Blood (19), was provided by R. Gallo (National Cancer Institute, Bethesda, MD). Cell lines were maintained in suspension in RPMI 1640 medium with 2 g of sodium bicarbonate per liter, 1% penicillin/streptomycin and 1% L-glutamine stock solutions, and 10% heat-inactivated fetal calf serum in Coming TM 25 flasks. Defined media were prepared as described (11). Doubling times were estimated only when cultures contained more than 98% viable cells as determined by trypan blue exclusion.Long-Term Human Bone Marrow Culture and ColonyForming Cell Ass...

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“…Serum free culture was carried out by using a modification of the technique described previously (20,21) . Culture mixture contained 1% crystallized globulin-free deionized BSA (Calbiochem-Behring Corp ., La Jolla, CA), 300 hg/ml fully saturated human transferrin (Sigma Chemical Co .…”

Section: Methodsmentioning

confidence: 99%

Stimulation of connective tissue-type mast cell proliferation by crosslinking of cell-bound IgE.

Takagi

Nakahata

Koike

et al. 1989

The Journal of Experimental Medicine

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Crosslinking of cell-bound IgE on mouse connective tissue-type mast cells (CTMC) by multivalent antigen or anti-IgE antibody induced clonal growth of CTMC in methylcellulose culture containing IL-3. Continuous presence of antigen, IgE antibody, and IL-3 in culture was required for extensive proliferation of CTMC. Optimal concentrations of antigen and anti-IgE antibody for proliferation of sensitized CTMC approximately corresponded to those for maximal histamine release from the cells, and it was observed that most dividing cells stimulated by antigen had pericellular degranulation halos in culture. Experiments of both single cell culture and serum free culture provided evidence for a direct effect of antigen stimulation on proliferation of CTMC. Neither accessory cells nor some factors in FCS were required for the clonal growth of CTMC in our culture condition. Compound 48/80, a direct stimulator of CTMC, also triggered histamine release from CTMC but failed to support their proliferation. These results suggest that stimulation of CTMC via IgE receptors not only triggers the release of chemical mediators from the cells but induces clonal growth of CTMC in the presence of IL-3. Our data indicate the possibility that antigen stimulation may play another role in the proliferation of CTMC.

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Complete replacement of serum in primary cultures of erythropoietin-dependent red cell precursors (CFU-E) by albumin, transferrin, iron, unsaturated fatty acid, lecithin and cholesterol

Cited by 243 publications

References 13 publications

A new candidate for the regulation of erythropoiesis

A new candidate for the regulation of erythropoiesis

Fibrinogen and its fragment D stimulate proliferation of human hemopoietic cells in vitro.

Stimulation of connective tissue-type mast cell proliferation by crosslinking of cell-bound IgE.

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