Eosinophil colony formation in semisolid cultures of human bone marrow cells

Abstract

A method for in situ staining and scoring of eosinophil colonies and clusters in the whole culture dish has been developed for agar cultures of human marrow cells. Cultures stimulated by human placental conditioned medium were found to develop the same proportion of eosinophil, neutrophil, and monocyte colonies as cultures stimulated by conventional peripheral white cell underlayers. Eosinophil colonies and clusters always contained pure populations of eosinophils, and no mixed colonies of eosinophils with other cells were observed. Eosinophil colony formation was delayed in onset relative to neutrophil or monocyte colony formation, but by day 14 eosinophil colonies comprised 20% of all colonies. Eosinophil colonies were absent in cultures of marrow cells from patients with acute myeloid leukemia in relapse, but eosinophil colonies and clusters were found with higher than normal frequency in cultures from acute leukemic patients in remission. A high frequency of eosinophil colonies and clusters was also observed in cultures from three patients with idiopathic thrombocytopenic purpura. The new staining procedure should permit a wide range of studies on eosinophil precursor populations in the marrows of patients with various diseases.

Separate actions of different colony stimulating factors from human placental conditioned medium on human hemopoietic progenitor cell survival and proliferation

“…Conversely, colonies initiated by CSFP degenerated during the second week of culture and this fate was not significantly influenced by substituting CSFas the stimulus. Previous studies (Dresch et al, 1979;Johnson et al, 1977) indicated that these subsets of GM progenitors could be partially separated on the basis of different sedimentation velocities and the present studies documented that a similar partial segregation could be achieved by FACS separation using the fact that day 7 colony-forming cells bound more fucose-binding protein than day 14 colonyforming cells. Unfortunately only a few eosino-phi1 clusters were transferred.…”

“…There is a heterogeneity in the physical characteristics of human colony-forming cells stimulated by human placental-conditioned medium based on whether they produce granulocyte, macrophage, mixed granulocyte-macrophage, or eosinophil colonies (Dresch et al, 1977). There are also two types of colonyforming cells based on velocity sedimentation studies.…”