The production of differentiation autoinducing activity by WEHI-3B D+ leukemia cells.
We studied the differentiation autoinducing activity in WEHI-3B D+ cell-conditioned medium (WCM). After culturing 10(6)/ml WEHI-3B D+ cells in RPMI-1640 medium without fetal calf serum (FCS) for 4 days, the supernatant was collected. The medium, concentrated 50-fold by YM-5 membrane filtration, was fractionated by gel exclusion on Ultrogel AcA44. We evaluated the effect of each of the four fractions on differentiation in WEHI-3B D+ cells by morphological, functional, and cytochemical criteria after adding the fractions to liquid or soft-agar cultures of 10(3) cells in 1 ml RPMI-1640 medium containing 10% FCS; the experimental cultures contained 10% of the fractions, with a control for each without the fraction. The growth of WEHI-3B D- cells in culture was inhibited by the addition of fraction P only (mol. wt. 10,000-20,000 daltons). In these same cultures, the cells were granulocyte-like, strongly positive for naphthol ASD chloroacetate esterase, and had phagocytic activity. Colonies grown in agar culture with fraction P also exhibited a peripheral halo of loosely dispersed cells around a central aggregate. Fraction P contained neither granulocyte colony-stimulating activity nor burst-promoting activity. These results suggest that fraction P contains differentiation autoinducing factor that is different from granulocyte colony-stimulating factor or interleukin 3.
Kajigaya
Y
Y
Department of Pediatrics, Yokohama City University School of Medicine, Japan.
Ikuta
K
K
Sasaki
H
H
Funabiki
T
T
Koiso
Y
Y
Matsuyama
S
S
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D005591:Chemical Fractionation; D003115:Colony-Stimulating Factors / Q000502:physiology; D003470:Culture Media / Q000032:analysis*; D004920:Erythropoiesis / Q000187:drug effects; D005260:Female; D006133:Growth Substances / Q000502:physiology*; D036341:Intercellular Signaling Peptides and Proteins; D007377:Interleukin-3; D007951:Leukemia, Myeloid / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology*
D003115:Colony-Stimulating Factors; D003470:Culture Media; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D007377:Interleukin-3; C047526:leukemia growth-promoting factor