International Journal of Cell Cloning, Vol 6, 281-289, Copyright © 1988 by AlphaMed Press
Hematopoiesis on cellulose ester membranes (CEM). IX. Enrichment of membranes with adherent layers from long-term marrow culture from normal or drug-treated mice (hematopoietic microenvironment study II)
SS Adler, SG Husseini and WH Knospe
Department of Medicine, Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612.
Cellulose ester membranes (Millipore) or polytetrafluoroethylene (Mitex)
membranes were coated with adherent layers taken from Dexter- type
long-term cultures, 4-5, 8 or 12 weeks after initiation of culture. The
cultures were established with marrow taken from untreated mice or, in some
cases, from mice treated with a single lethal dose (LD10) of carmustine
(BCNU) or cyclophosphamide. In the studies using untreated mice, the
cultures went for 8 or 12 weeks and in the drug studies, for 4-5 weeks. The
8 and 12 week cultures were reseeded at 4 weeks. The membranes were
implanted into the peritoneal cavities of mice for 3-12 months after which
they were removed, fixed, sectioned and stained for histologic study. After
6 months of implantation, about 40% of the membranes coated with cells from
non-drug-treated mice and 60% of the membranes coated with cells from
drug-treated mice contained hematopoietic elements; often there were foci
of trilineal hematopoiesis. Hematopoiesis never occurred without bone
formation, but the reverse was not true. Membranes coated with adherent
layers established from marrow of mice treated with cyclophosphamide or
BCNU showed two main characteristics: 1) they supported hematopoiesis
normally, and 2) the regeneration of stroma and hematopoiesis occurred
earlier than in membranes coated with stroma derived from normal mice,
perhaps because the cells from the drug-treated mice spent a shorter time
in culture. In vitro culture may damage cells required to condition the
membrane for hematopoiesis.
