International Journal of Cell Cloning, Vol 10, 33-46, Copyright © 1992 by AlphaMed Press
Isolation of permanent clonal bone marrow stromal cell lines derived from "viable moth-eaten" and "severe combined immunodeficiency" mutant mice
A Ohara, P Anklesaria, L Shultz and JS Greenberger
Department of Radiation Oncology, University of Massachusetts Medical Center, Worcester 01655.
Long-term bone marrow cultures (LTBMCs) were established from bone marrow
of mev/mev mice with severe combined immunodeficiency (scid) and their
normal littermates, and permanent stromal cell lines were derived. In
cultures from mutant mev/mev mice, an adherent layer containing around 10%
of fibro-endothelial cells and 90% macrophage- like cells was observed. A
permanent cell line actively inhibited hematopoiesis resulting in an
80-fold decrease in production of cumulative colony-forming unit culture
units (CFU) including granulocyte, erythroid, macrophage, megakaryocyte
(CFU-GEMM) and granulocyte-macrophage (CFU-GM). To confirm active
inhibition, we performed coculture experiments using permanent cell lines
and engrafted each in vitro with hematopoietic progenitors from normal
mouse LTBMCs. "Cobblestone island" and nonadherent cell production were
observed adding hemopoietic progenitors to coculture of fibroblastic cells
of mev/mev cultures with lines from normal littermates. In contrast,
macrophage cell line Mev #5 suppressed hemopoiesis and itself had a
proliferative advantage. Thus, bone marrow stromal cell lines from mev/mev
mice reproduce some characteristics of the hematopoietic microenvironmental
defect in vitro.
