Support of human cord blood progenitor cells on human stromal cell lines
transformed by SV40 large T antigen under the influence of an inducible
(metallothionein) promoter
FM Cicuttini, M Martin, E Salvaris, L Ashman, CG Begley, J Novotny, D Maher and AW Boyd
Lions Clinical Cancer Research Laboratory, Walter and Eliza Hall Institute
of Medical Research, PO Royal Melbourne Hospital, Victoria, Australia.
We describe the development of a human bone marrow (BM) culture system
which allows study of the interaction of stromal cell lines (SCL) and
highly purified hematopoietic progenitor cells. Normal BM stromal cells
were electroporated with a plasmid containing the simian virus 40 (SV40)
large T antigen (SV40 T Ag) under the control of a synthetic
metallothionein promoter (MT4); this construct is designated MT4 SV40 T Ag.
SCL in which the rate of proliferation could be controlled by altering the
zinc (Zn) concentration were characterized, demonstrating that the SCL were
heterogeneous with respect to G-CSF and GM-CSF production. Suppression of
SCL proliferation on removal of Zn made it possible to use these lines in
coculture with purified CD34+ progenitor cells from umbilical cord blood.
The ability to control proliferation of SCL has allowed us to maintain the
survival and expansion of colony- forming cells in culture for up to 2
months. These lines have enabled us to test for stromal cell
characteristics at a clonal level and provided us with a tool to analyze
the events leading to lineage commitment and hematopoietic differentiation,
as demonstrated by suppression of hematopoiesis by an antibody directed
against the c-kit molecule.
Volume 80,
Issue 1,
pp. 102-112,
07/01/1992
Copyright © 1992 by The American Society of Hematology
