Isolation and biological characterization of two classes of blast-cell
colony-forming cells from normal murine marrow
G Migliaccio, M Baiocchi, JW Adamson and AR Migliaccio
Laboratory of Hematopoietic Growth Factors, Lindsley F. Kimball Research
Institute, New York Blood Center, NY 10021, USA.
In this study, a primitive progenitor cell, the blast-cell colony- forming
cell (BC-CFC), which is thought by some to be equivalent to the
hematopoietic stem cell (HSC), those cells capable of long-term marrow
repopulation, has been isolated from normal murine marrow. The cell
separation method we employed has, as its final step, the purification of
marrow cells based on their ability to take up (bright) or exclude (dull)
the mitochondrial dye, Rhodamine (Rho)-123. Rho-bright and Rho- dull cells
are enriched for multipotential progenitor cells or for HSC, respectively.
It was found that Rho-bright cells contain more BC-CFC than Rho-dull cells
(310 +/- 50 v 120 +/- 40 per 10(5) purified cells, respectively). However,
the BC-CFC that copurified with the Rho-bright and the Rho-dull cells were
different in terms of replating efficiency and response to interleukin-3
(IL-3) and stem cell factor (SCF). In fact, on replating, the blast-cell
colonies cultured from the Rho-dull population give rise to many more
secondary colonies and had a greater replating efficiency than those
obtained from Rho-bright cells (replating efficiency: 29.0 +/- 6.3 v 19.5
+/- 3.7, respectively, P < .01). Furthermore, while the same numbers of
blast-cell colonies were detected in culture of Rho-bright cells stimulated
with IL-3 alone or in combination with SCF, blast-cell colonies were
generated in cultures of Rho-dull cells only in the presence of both IL-3
and SCF. After 5 days in suspension culture stimulated with IL-3 and SCF,
Rho-dull cells generated BC-CFC whose replating potential was similar to
the replating potential of BC-CFC contained in the Rho-bright population.
These results indicate that BC-CFC contained in the Rho-bright and -dull
populations are qualitatively different. Because the Rho-dull population
contains HSC, the results indicate that few, if any, BC-CFC are HSC.
Volume 87,
Issue 10,
pp. 4091-4099,
05/15/1996
Copyright © 1996 by The American Society of Hematology
