title: Promotion of survival and proliferation by interleukin 3, kit-ligand and erythropoietin on early and late appearing spleen colony forming units in culture.
abstract: We examined the effects of interleukin 3 (IL-3), kit-ligand and erythropoietin (EPO) on the survival and growth of early appearing spleen colony forming units (CFU-S8) and late appearing CFU-S (CFU-S12) in short-term liquid culture (SLC). In the control cultures, without any additive, CFU-S8 and CFU-S12 declined; nearly 10% of the initial number of CFU-S still survived by the second day of culture. The addition of IL-3 or kit-ligand increased the survival both of CFU-S8 and CFU-S12, with an increased dose of concentration, at final concentrations of 10-200 U/ml and 500-50x dilution, respectively. However, only the survival of CFU-S8 increased when EPO was added up to 10 U/ml, while the frequency of CFU-S12 was not higher than in the control culture. The percentage of CFU-S8 and CFU-S12 in DNA synthesis, evaluated in 3H-TdR cytocide experiments, increased after one day in culture with each of the three factors. The results suggest that all three factors stimulated the proliferation of both populations of CFU-S, but the two populations showed different patterns of response to each factor; EPO stimulates the proliferation of CFU-S12 that differentiate into CFU-S8 that have less capacity for self-renewal, while the addition of IL-3 or kit-ligand causes an increase in the number of both populations due to the stimulation of an earlier stage of stem cell differentiation or self-renewal of CFU-S12. Our experimental system (SLC-CFU-S assay) is useful for evaluating the response of hematopoietic stem cells to cytokines which promote the in vitro survival and proliferation of these cells.
authors.affiliation: Department of Pediatrics, Yokohama City University, School of Medicine, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Sasaki
authors.forename: Y
authors.initials: Y
authors.lastname: Hirabayashi
authors.forename: T
authors.initials: T
authors.lastname: Inoue
authors.forename: S
authors.initials: S
authors.lastname: Taniguchi
authors.forename: K
authors.initials: K
authors.lastname: Ikuta
authors.forename: S
authors.initials: S
authors.lastname: Matsuyama
mesh_terms: D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D004261:DNA Replication / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D007377:Interleukin-3 / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011994:Recombinant Proteins / Q000494:pharmacology; D013154:Spleen / Q000166:cytology*; D019089:Stem Cell Factor
chemical_list: D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D004921:Erythropoietin