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Culture of phenotypically defined hematopoietic stem cells and other
progenitors at limiting dilution on Dexter monolayers [see comments]
K Weilbaecher, I Weissman, K Blume and S Heimfeld
SyStemix, Palo Alto, CA.
Highly enriched, phenotypically defined hematopoietic stem, Thy-1loLin-
Sca-1+, and progenitor cell populations from mouse bone marrow (BM) were
tested at limiting dilution for their ability to reconstitute Dexter
monolayers. Several classes of BM cells can reconstitute Dexter cultures,
first forming discrete "cobblestone" areas which then mature into colonies
consisting primarily of maturing myeloid and erythroid cells. Most such
colonies have a limited lifespan in culture. Only the Thy-1loLin-Sca-1+
cell fraction gives rise to colonies that survive longer than 3 weeks,
which suggests that a limiting-dilution analysis for long-term
reconstitution of Dexter cultures can serve as a quantitative measure of
stem cell activity. Additional experiments were performed to assess the
formation of new progenitor cells in reconstituted Dexter cultures. Again,
only cultures seeded with the stem cell-enriched fraction contained
expanded numbers of replatable WEHI-3 CM responsive colony-forming cells
(CFU-GM). Quantitative analysis indicates that 97% of the replatable CFU-GM
of whole BM is contributed by the Thy-1loLin-Sca-1+ cell fraction, again
suggesting a potential stem cell-specific assay. Such quantitative in vitro
assays might prove useful in characterization and isolation of human stem
cells where in vivo assays are lacking.
Volume 78,
Issue 4,
pp. 945-952,
08/15/1991
Copyright © 1991 by The American Society of Hematology
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