Growth characteristics of bone marrow cells from beige mutant, the mouse homologue of the Chediak-Higashi syndrome of man, propagated in semisolid agar cultures.

Abstract

Suspensions of bone marrow cells from the beige (bg/bg) mouse, a homologue of the Chediak-Higashi syndrome (C-HS) of man, and normal mouse bone marrow cells, when stimulated by colony-stimu.ating factor (CSF) from different sources, proliferate in semisolid agar cultures and produce colonies composed of granulocytic and/or mononuclear cells. Studies with CSF from various sources (embryo and kidney feeder monolayers, conditioned media from embryo and kidney cell cultures, and plasma from untreated, irradiated, or endotoxin-treated mice) indicated that bone marrow cells from the beige mouse are quantitatively as capable of developing into colonies as normal cells when incubated with the same CSF source. Experiments which compared the bone marrow colony response of beige and normal cells using postendotoxin plasma (PEP) as the CSF source indicated that beige cells were stimulated to the same extent by tcsf obtained from either the normal or the beige mouse. CSF obtained from normal or affected mice was equally effective in stimulating bone marrow cell proliferation. There was no discordance of colony cell types present when PEP-stimulated 8-day bone marrow colonies from normal and affected mice were compared. The nuclear morphology of beige and normal granulocytes from these cultures indicated cells ranging from myeloblasts to mature segmented polymorphonuclear leukocytes. These preliminary studies indicate that the agar culture method for the growth of mononuclear and granulocytic colonies may provide a method for obtaining enriched C-HS cell populations for biochemical analysis of the genetic defect(s) in this interesting disease.