Growth and differentiation of a murine interleukin-3-producing myelomonocytic leukemia cell line in a protein-free chemically defined medium.
We established the continuous growth of WEHI-3B D+ cells in protein-free chemically defined F-12 medium by stepwise decreases in the concentration of fetal calf serum. This cell line, designated as WEHI-3B-Y1, has now been propagated in protein-free F-12 medium for 3 years. The population-doubling time of the cells in culture is about 24 hr. WEHI-3B-Y1 cells are immature undifferentiated cells which show positive staining for naphthol ASD chloroacetate esterase and alpha-naphthyl butyrate esterase and spontaneously exhibit a low level of differentiation to mature granulocytes and macrophages. Medium conditioned by WEHI-3B-Y1 cells stimulated the proliferation of an interleukin-3 (IL-3)-dependent FDCP-2 cell line. This conditioned medium was shown to have erythroid burst-promoting activity when assayed using normal murine bone marrow. The colony formation of WEHI-3B-Y1 cells in semi-solid agar culture was not stimulated by purified recombinant human granulocyte colony-stimulating factor (rhG-CSF). However, in the presence of human transferrin, rhG-CSF enhanced the number of colonies of WEHI-3B-Y1 cells but did not induce their differentiation. These results suggest that WEHI-3B-Y1 cells cultured in protein-free medium produced murine IL-3. In addition, human G-CSF enhanced the clonal growth but did not induce the differentiation of WEHI-3B-Y1 cells cultured in serum-free medium.
Kajigaya
Y
Y
Department of Pediatrics, Yokohama City University School of Medicine, Japan.
Ikuta
K
K
Sasaki
H
H
Matsuyama
S
S
D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D003470:Culture Media; D015672:Erythroid Precursor Cells / Q000473:pathology; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D007377:Interleukin-3 / Q000096:biosynthesis*; D015479:Leukemia, Myelomonocytic, Acute / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011506:Proteins / Q000008:administration & dosage; D011994:Recombinant Proteins / Q000494:pharmacology; D014168:Transferrin / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology; D014410:Tumor Stem Cell Assay
D003470:Culture Media; D007377:Interleukin-3; D011506:Proteins; D011994:Recombinant Proteins; D014168:Transferrin; D016179:Granulocyte Colony-Stimulating Factor