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 Previous Article | Table of Contents | Next Article 
Enrichment and characterization of murine hematopoietic stem cells that
express c-kit molecule
S Okada, H Nakauchi, K Nagayoshi, S Nishikawa, S Nishikawa, Y Miura and T Suda
Department of Medicine, Jichi Medical School, Tochigi-ken, Japan.
The proto-oncogene c-kit encodes a transmembrane tyrosine kinase receptor
for stem cell factor (SCF). The c-kit/SCF signal is expected to have an
important role in hematopoiesis. A monoclonal antibody (ACK- 2) against the
murine c-kit molecule was prepared. Flow cytometric analysis showed that
the bone marrow cells that expressed the c-kit molecule (approximately 5%)
were B220(B)-, TER119(erythroid)-, Thy1negative-low, and WGA+. A small
number of Mac-1(macrophage)+ or Gr- 1(granulocyte)+ cells were c-kit-low
positive. Colony-forming unit in culture (CFU-C) and day-8 and day-12
CFU-spleen (CFU-S) existed exclusively in the c-kit-positive fraction.
About 20% of the Lin(lineage)-c-kit+ cells were rhodamine-123low and this
fraction contained more day-12 CFU-S than day-8 CFU-S. On the basis of
these findings, murine hematopoietic stem cells were enriched with normal
bone marrow cells. One of two and one of four Thy-1lowLin-WGA+c-kit+ cells
were CFU-C and CFU-S, respectively. Long-term repopulating ability was
investigated using B6/Ly5 congenic mice. Eight and 25 weeks after
transplantation of Lin-c-kit+ cells, donor-derived cells were found in the
bone marrow, spleen, thymus, and peripheral blood. In peripheral blood, T
cells, B cells, and granulocyte-macrophages were derived from donor cells.
Injection of ACK-2 into the irradiated mice after bone marrow
transplantation decreased the numbers of day-8 and day-12 CFU-S in a
dose-dependent manner. Day-8 spleen colony formation was completely
suppressed by the injection of 100 micrograms ACK-2, but a small number of
day-12 colonies were spared. Our data show that the c- kit molecule is
expressed in primitive stem cells and plays an essential role in the early
stages of hematopoiesis.
Volume 78,
Issue 7,
pp. 1706-1712,
10/01/1991
Copyright © 1991 by The American Society of Hematology
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