Complete replacement of serum in cultures of murine primitive erythroid and multipotential progenitor cells: absolute requirement for spleen conditioned medium.
We describe a serum-free medium for the formation of erythropoietic bursts by murine bone marrow cells. Iscove's modified Dulbecco's medium supplemented with bovine serum albumin, iron-saturated transferrin, soybean phospholipids and cholesterol supported burst formation. The further addition of hemin increased burst numbers to above those obtained in serum-containing cultures. With or without hemin, a source of burst-promoting activity (BPA) (crude or partially purified spleen conditioned medium) and erythropoietin were essential. This system provides a sensitive assay for BPA. Of all colonies developing in these cultures, 16% were pure erythroid, 17% mixed erythroid/myeloid, 36% macrophage, 19% macrophage/basophil and macrophage/neutrophil, 9% basophil and 2% neutrophil.
D000818:Animals; D001769:Blood; D001854:Bone Marrow Cells; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media; D004920:Erythropoiesis; D004921:Erythropoietin / Q000494:pharmacology; D005260:Female; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006427:Hemin / Q000494:pharmacology; D066298:In Vitro Techniques; D008055:Lipids / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013154:Spleen / Q000166:cytology; D014168:Transferrin / Q000494:pharmacology
D003470:Culture Media; D008055:Lipids; D014168:Transferrin; D004921:Erythropoietin; D012710:Serum Albumin, Bovine; D006427:Hemin