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Thrombopoietin Promotes the Survival of Murine Hematopoietic
Long-Term Reconstituting Cells: Comparison With the Effects of
FLT3/FLK-2 Ligand and Interleukin-6
Takuya Matsunaga,
Takashi Kato,
Hiroshi Miyazaki, and
Makio Ogawa
From the Department of Veterans Affairs Medical Center and the
Department of Medicine, Medical University of South Carolina,
Charleston, SC; and the Pharmaceutical Research Laboratory, Kirin
Brewery Co, Ltd, Takasaki, Japan.
The effects of thrombopoietin (TPO; c-mpl ligand),
FLT3/FLK-2 ligand (FL), and interleukin-6 (IL-6) on the survival of
murine hematopoietic long-term reconstituting cells (LTRC) were studied by using lineage-negative, Sca-1-positive, c-kit-positive
(Lin Sca-1+c-kit+) marrow
cells from 5-fluorouracil-treated mice. We tested the ability of these
cytokines to maintain the viability of LTRC by transplanting the
cultured cells to lethally irradiated Ly-5 congenic mice together with
compromised marrow cells. As a single agent, only TPO could maintain
the LTRC. Neither IL-6 nor FL was effective by itself, but they acted
synergistically to maintain the LTRC. We examined whether the
maintenance of LTRC by these cytokines was due to the survival of stem
cells or was the result of active cell divisions and self-renewal. To
monitor cell division, we used membrane dye PKH26. Enriched cells were
stained with PKH26 on day 0 and incubated in suspension culture with
TPO or with IL-6 and FL for 7 days. On day 7, PKH26low and
PKH26high cells were prepared by sorting and their in vivo
reconstituting abilities were tested by transplantation into lethally
irradiated Ly-5 congenic mice together with compromised marrow cells.
PKH26high populations cultured with both TPO alone and the
combination of IL-6 and FL showed greater reconstitution activity than
that of PKH26low populations. These data indicate that TPO
alone and the combination of IL-6 and FL can support the survival of
stem cells without stimulating their active cell proliferation.
Blood, Vol. 92 No. 2 (July 15), 1998:
pp. 452-461
© 1998 by the American Society of Hematology.
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