Two classes of primitive pluripotent hemopoietic progenitor cells: separation by adherence.
Methylcellulose cultures containing mouse marrow cells at low densities and partially purified preparations of erythropoietin and Interleukin-3 were scored after 2 weeks for the presence of macroscopic multilineage colonies (from "primary" CFU-macro GEMM). Whole cultures were then harvested and replanted to assess the number of "secondary" CFU-macro GEMM produced, but not detected, during the primary culture period. In such experiments adherent marrow cells yielded significantly higher numbers of secondary CFU-macro GEMM than did either fresh or nonadherent marrow cells. Removal of macroscopic colonies prior to replating showed that most secondary CFU-macro GEMM were not derived from primary CFU-macro GEMM. In vivo studies also revealed a differential effect of adherence separation on the frequency of day 10 CFU-S, which decreased, by comparison to cells capable of long-term repopulation, which increased. Primary adherent CFU-macro GEMM from 5-fluorouracil (5-FU) treated mice showed an 18-fold higher self-renewal capacity than their counterparts in normal marrow. Nevertheless the majority of secondary CFU-macro GEMM obtained from primary cultures of adherent 5-FU cells were again not derived from primary CFU-macro GEMM. Cells capable of immediately generating large multilineage colonies thus appear to represent an intermediate compartment of pluripotent progenitors whose self-renewal properties, may, however, vary over a considerable range. Our results further suggest that these progenitors are derived ultimately from a more primitive adherent cell whose tendency to begin to divide in vitro is low and whose presence correlates with cells capable of long-term myeloid repopulation in vivo.
D000818:Animals; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002469:Cell Separation; D003114:Colony-Forming Units Assay; D005472:Fluorouracil / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008747:Methylcellulose; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL
D008747:Methylcellulose; D005472:Fluorouracil