|
|
 Previous Article | Table of Contents | Next Article 
Stem cell factor influences the proliferation and erythroid differentiation
of the MB-02 human erythroleukemia cell line by binding to a high-affinity
c-kit receptor
VC Broudy, DA Morgan, N Lin, KM Zsebo, FW Jacobsen and T Papayannopoulou
Department of Medicine, University of Washington, Seattle 98195.
Stem cell factor (SCF) acts in synergy with other growth factors such as
erythropoietin (Epo), granulocyte-macrophage colony-stimulating factor
(GM-CSF), or interleukin-3 (IL-3), to stimulate the growth of primitive
hematopoietic cells. Because of the prominent role of CSF in the
maintenance of normal erythropoiesis in vivo, we examined the effects of
SCF on the Epo-inducible human erythroleukemia cell line MB- 02, and
characterized the c-kit receptor in these cells. MB-02 cells cultured in
serum-containing media do not survive in the absence of exogenous growth
factors, but the addition of SCF, Epo, or IL-3 as a single factor enhanced
MB-02 survival. Furthermore, in the presence of Epo, SCF (5 to 25 ng/mL)
enhanced MB-02 proliferation in a dose- dependent manner, and increased the
relative and absolute number of benzidine-positive cells generated. SCF
also enhanced cell proliferation in the presence of either IL-3 or low
concentrations of GM-CSF. A neutralizing anti-c-kit receptor monoclonal
antibody (SR-1) blocked binding of 125I-SCF to MB-02 cells by 98%, and the
effect of SCF on MB-02 growth, c-kit receptor-binding parameters were
quantitated by equilibrium-binding experiments with 125I-SCF. MB-02 cells
display a single class of high-affinity (50 pmol/L) c-kit receptors, with
approximately 8,000 receptors per cell. The molecular weight of the c- kit
receptor was determined by affinity cross-linking 125I-SCF to MB-02 cells.
125I-SCF-c-kit receptor complexes of approximately 155,000 and
approximately 310,000 daltons were found, likely representing the monomeric
and dimeric forms of the c-kit receptor. The binding affinity and molecular
weight of the c-kit receptor on MB-02 cells are similar to those of normal
human marrow cells. These results suggest that SCF synergizes with Epo to
influence not only the proliferation but the erythroid differentiation of
MB-02 cells. Thus, the MB-02 cell line may be a useful model in which to
investigate the molecular mechanisms of SCF action.
Volume 82,
Issue 2,
pp. 436-444,
07/15/1993
Copyright © 1993 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
S. Ito, C. R. Mantel, M.-K. Han, S. Basu, S. Fukuda, S. Cooper, and H. E. Broxmeyer
Mad2 is required for optimal hematopoiesis: Mad2 associates with c-Kit in MO7e cells
Blood,
March 1, 2007;
109(5):
1923 - 1930.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Ghose, H. Porzig, and K. Baltensperger
Induction of Erythroid Differentiation by Altered Galpha 16 Activity as Detected by a Reporter Gene Assay in MB-02 Cells
J. Biol. Chem.,
April 30, 1999;
274(18):
12848 - 12854.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. C. Broudy, N. L. Lin, H.-J. Buhring, N. Komatsu, and T. J. Kavanagh
Analysis of c-kit Receptor Dimerization by Fluorescence Resonance Energy Transfer
Blood,
February 1, 1998;
91(3):
898 - 906.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Joneja, H.-C. Chen, D. Seshasayee, A. L. Wrentmore, and D. M. Wojchowski
Mechanisms of Stem Cell Factor and Erythropoietin Proliferative Co-signaling in FDC2-ER Cells
Blood,
November 1, 1997;
90(9):
3533 - 3545.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Sakatoku and S. Inoue
In Vitro Proliferation and Differentiation of Erythroid Progenitors of Cord Blood
Stem Cells,
July 1, 1997;
15(4):
268 - 274.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
V. C. Broudy, N. L. Lin, D. F. Sabath, T. Papayannopoulou, and K. Kaushansky
Human Platelets Display High-Affinity Receptors for Thrombopoietin
Blood,
March 15, 1997;
89(6):
1896 - 1904.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
