Korean J Hematol 2001; 36(4):
Published online December 31, 2001
© The Korean Society of Hematology
조덕연, 황진희, 곽승근, 신현영, 김성은, 윤환중, 김삼용, 성주명
충남대학교 의과대학 내과학교,
이화여자대학교 의과대학 내과학교실
Background: There has been contradictory reports regarding the homing potential of hematopoietic cells briefly exposed to hematopoietic growth factors in vitro. To get a resolution to this controversy, we investigated the effects of
short-term growth factor treatment of hematopoietic cells on the expression of CXCR4 and adhesion molecules, and the chemotaxis in response to stromal cell-dervied factor-1 (SDF-1), which is widely accepted to play a critical role in bone marrow (BM) homing of hematopoietic stem cells.
Methods:
BM and cord blood(CB) CD34+ cells were incubated with various hematopoietic growth factors including IL-1β, IL-3, IL-6, G-CSF, GM-CSF, stem cell factor (SCF), flk-2 ligand, and thrombopoietin, alone or in combination for up to 48 hours. Before and after the incubation, the expression of CXCR4 and adhesion molecules of CD34+ cells was analyzed using flow cytometry. SDF-1-mediated transmembrane or transendothelial migration of CD34+ cells,
cobblestone area-forming cells CD34+ cells, cobblestone area-forming cells (CAFCs), and/or long-term culture-initiating cells (LTC-ICs) was measured using Transwell M system.
Results:
VLA-4 was moderately up-refulated by the incubation of
the cells with IL-3 and SCF, and ICAM-1 was slightly up-regulated by IL-1β and IL-3. The expression of L-selectin, PECAM-1 or LFA-1 was not altered by any growth factors. With the incubation of the cells in the absence of growth factors or SDF-1, CXCR4 expression of CD34+ cells was rapidly increased, reaching a plateau at 24 hours. The spontaneous up-regulation was abrogated with the addition of SDF-1. In agreement with the up-regulation of CXCR4, CD34+ cells incubated for 40 hours showed much enhanced chemotaxis in response to SDF-1 compared to non-incubated cells (24.7±3.5% vs. 7.0±1.6%, P=0.01). Any growth factors examined in this study did not alter the CXCR4 expression of CD34+ cells. Neither did growth factors affect the transendothelial migration of LTC-ICs toward bone marrow stromal cells as well as the SDF-1-induced transmembrane chemotaxis of CD34+ cells and CAFCs.
Conclusion:
Short-term treatment of hemotopoietic cells with hematopoietic growth factors does not alter the expression of CXCR4 or SDF-1-mdeiated transendothelial chemotaxis.
Keywords Hematopoietic cells, CXCR4, SDF-1, Adhesion molecules, Bone marrow homing, Hematopoietic growth factors
Korean J Hematol 2001; 36(4): 324-334
Published online December 31, 2001
Copyright © The Korean Society of Hematology.
조덕연, 황진희, 곽승근, 신현영, 김성은, 윤환중, 김삼용, 성주명
충남대학교 의과대학 내과학교,
이화여자대학교 의과대학 내과학교실
Deog Yeon Jo, Jin Hee Hwang, Seung Keun Kwak, Hyun Young Shin, Sung Eun kim, Hwan Jung Yun, Sam Yong Kim, Chu Myong Seong
Department of Internal Medicine, College of Medicine, Chungnam National University, Taejon
Ewha Womans University, Seoul, Korea
Background: There has been contradictory reports regarding the homing potential of hematopoietic cells briefly exposed to hematopoietic growth factors in vitro. To get a resolution to this controversy, we investigated the effects of
short-term growth factor treatment of hematopoietic cells on the expression of CXCR4 and adhesion molecules, and the chemotaxis in response to stromal cell-dervied factor-1 (SDF-1), which is widely accepted to play a critical role in bone marrow (BM) homing of hematopoietic stem cells.
Methods:
BM and cord blood(CB) CD34+ cells were incubated with various hematopoietic growth factors including IL-1β, IL-3, IL-6, G-CSF, GM-CSF, stem cell factor (SCF), flk-2 ligand, and thrombopoietin, alone or in combination for up to 48 hours. Before and after the incubation, the expression of CXCR4 and adhesion molecules of CD34+ cells was analyzed using flow cytometry. SDF-1-mediated transmembrane or transendothelial migration of CD34+ cells,
cobblestone area-forming cells CD34+ cells, cobblestone area-forming cells (CAFCs), and/or long-term culture-initiating cells (LTC-ICs) was measured using Transwell M system.
Results:
VLA-4 was moderately up-refulated by the incubation of
the cells with IL-3 and SCF, and ICAM-1 was slightly up-regulated by IL-1β and IL-3. The expression of L-selectin, PECAM-1 or LFA-1 was not altered by any growth factors. With the incubation of the cells in the absence of growth factors or SDF-1, CXCR4 expression of CD34+ cells was rapidly increased, reaching a plateau at 24 hours. The spontaneous up-regulation was abrogated with the addition of SDF-1. In agreement with the up-regulation of CXCR4, CD34+ cells incubated for 40 hours showed much enhanced chemotaxis in response to SDF-1 compared to non-incubated cells (24.7±3.5% vs. 7.0±1.6%, P=0.01). Any growth factors examined in this study did not alter the CXCR4 expression of CD34+ cells. Neither did growth factors affect the transendothelial migration of LTC-ICs toward bone marrow stromal cells as well as the SDF-1-induced transmembrane chemotaxis of CD34+ cells and CAFCs.
Conclusion:
Short-term treatment of hemotopoietic cells with hematopoietic growth factors does not alter the expression of CXCR4 or SDF-1-mdeiated transendothelial chemotaxis.
Keywords: Hematopoietic cells, CXCR4, SDF-1, Adhesion molecules, Bone marrow homing, Hematopoietic growth factors
Ha-Yon Kim, Ji-Young Hwang, Yoon-Suk Oh, Seong-Woo Kim, Hyo-Jin Lee, Hwan-Jung Yun, Samyong Kim, Young-Jun Yang, and Deog-Yeon Jo
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