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In vivo and in vitro stem cell function of c-kit- and Sca-1-positive murine
hematopoietic cells
S Okada, H Nakauchi, K Nagayoshi, S Nishikawa, Y Miura and T Suda
Department of Medicine, Jichi Medical School, Tochigi-ken, Japan.
c-kit is expressed on hematopoietic stem cells and progenitor cells, but
not on lymphohematopoietic differentiated cells. Lineage marker- negative,
c-kit-positive (Lin-c-kit+) bone marrow cells were fractionated by means of
Ly6A/E or Sca-1 expression. Lin-c-kit+Sca-1+ cells, which consisted of
0.08% of bone marrow nucleated cells, did not contain day-8 colony-forming
units-spleen (CFU-S), but 80% were day-12 CFU-S. One hundred cells rescued
the lethally irradiated mice and reconstituted hematopoiesis. On the other
hand, 2 x 10(3) of Lin-c- kit+Sca-1- cells formed 20 day-8 and 11 day-12
spleen colonies, but they could not rescue the lethally irradiated mice.
These data indicate that Lin-c-kit+Sca-1+ cells are primitive hematopoietic
stem cells and that Sca-1-cells do not contain stem cells that reconstitute
hematopoiesis. Lin-c-kit+Sca-1+ cells formed no colonies in the presence of
stem cell factor (SCF) or interleukin-6 (IL-6), and only 10% of them formed
colonies in the presence of IL-3. However, approximately 50% of them formed
large colonies in the presence of IL- 3, IL-6, and SCF. Moreover, when
single cells were deposited into culture medium by fluorescence-activated
cell sorter clone sorting system, 40% of them proliferated on a stromal
cell line (PA-6) and proliferated for more than 2 weeks. In contrast, 15%
of the Lin-c- kit+Sca-1-cells formed colonies in the presence of IL-3, but
no synergistic effects were observed in combination with SCF plus IL-6
and/or IL-3. Approximately 10% proliferated on PA-6, but most of them
degenerated within 2 weeks. The population ratio of c-kit+Sca-1+ to c-
kit+Sca-1- increased 2 and 4 days after exposure to 5-fluorouracil (5- FU).
These results are consistent with the relative enrichment of highly
proliferative colony-forming cells by 5-FU. These data show that, although
c-kit is found both on the primitive hematopoietic stem cells and
progenitors, Sca-1+ cells are more primitive and respond better than Sca-1-
cells to a combination of hematopoietic factors, including SCF and stromal
cells.
Volume 80,
Issue 12,
pp. 3044-3050,
12/15/1992
Copyright © 1992 by The American Society of Hematology
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