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Functionally distinct human marrow stromal cell lines immortalized by
transduction with the human papilloma virus E6/E7 genes
BA Roecklein and B Torok-Storb
Program in Transplantation Biology, Clinical Research Division, Fred
Hutchinson Cancer Research Center, Seattle, WA 98104.
A replication-defective recombinant retrovirus containing the human
papilloma virus E6/E7 genes (LXSN-16 E6E7) was used to immortalize stromal
cells from human marrow. The E6/E7 gene products interfere with the
function of tumor-suppressor proteins p53 and Rb, respectively, thereby
preventing cell cycle arrest without causing significant transformation.
Twenty-seven immortalized clones designated HS-1 to HS- 27 were isolated,
four of which are characterized in this report. Two cell lines, HS-5 and
HS-21, appear to be fibroblastoid and secrete significant levels of
granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-CSF
(GM-CSF), macrophage-CSF (M-CSF), Kit ligand (KL), macrophage-inhibitory
protein-1 alpha, interleukin-6 (IL-6), IL- 8, and IL-11. However, only HS-5
supports proliferation of hematopoietic progenitor cells when cocultured in
serum-deprived media with no exogenous factors. Conditioned media (CM) from
HS-5 promotes growth of myeloid colonies to significantly greater extent
than a cocktail of recombinant factors containing 10 ng/mL of IL-1, IL-3,
IL- 6, G-CSF, GM-CSF, and KL and 3 U of erythropoietin (Epo). Two
additional clones, HS-23 and HS-27, resemble "blanket" cells, with an
epithelioid morphology, and are much larger, broader, and flatter when
compared with HS-5 and HS-21. These lines secrete low levels of growth
factors and do not support proliferation of isolated progenitor cells in
cocultures. CM from HS-23 and HS-27 also fail to support growth of myeloid
colonies. Both HS-23 and HS-27 express relatively high levels of VCAM-1,
yet HS-27 is the only line that supports the formation of "cobblestone"
areas by isolated CD34+38lo cells. We hypothesize that HS- 5, HS-21, HS-23,
and HS-27 represent functionally distinct components of the marrow
microenvironment.
Volume 85,
Issue 4,
pp. 997-1005,
02/15/1995
Copyright © 1995 by The American Society of Hematology
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