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Chemotaxis of primitive hematopoietic cells in response to stromal cell–derived factor-1
Deog-Yeon Jo, … , Tsuneyoshi Hamada, Malcolm A.S. Moore
Deog-Yeon Jo, … , Tsuneyoshi Hamada, Malcolm A.S. Moore
Published January 1, 2000
Citation Information: J Clin Invest. 2000;105(1):101-111. https://doi.org/10.1172/JCI7954.
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Chemotaxis of primitive hematopoietic cells in response to stromal cell–derived factor-1

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Abstract

Stromal cell–derived factor-1 (SDF-1) provides a potent chemotactic stimulus for CD34+ hematopoietic cells. We cultured mobilized peripheral blood (PB) and umbilical cord blood (CB) for up to 5 weeks and examined the migratory activity of cobblestone area–forming cells (CAFCs) and long-term culture–initiating cells (LTC-ICs) in a transwell assay. In this system, SDF-1 or MS-5 marrow stromal cells placed in the lower chamber induced transmembrane and transendothelial migration by 2- and 5-week-old CAFCs and LTC-ICs in 3 hours. Transmigration was blocked by preincubation of input CD34+ cells with antibody to CXCR4. Transendothelial migration of CB CAFCs and LTC-ICs was higher than that of PB. We expanded CD34+ cells from CB in serum-free medium with thrombopoietin, flk-2 ligand, and c-kit ligand, with or without IL-3 and found that CAFCs cultured in the absence of IL-3 had a chemotactic response equivalent to noncultured cells, even after 5 weeks. However, addition of IL-3 to the culture reduced this response by 20–50%. These data indicate that SDF-1 induces chemotaxis of primitive hematopoietic cells signaling through CXCR4 and that the chemoattraction could be downmodulated by culture ex vivo.

Authors

Deog-Yeon Jo, Shahin Rafii, Tsuneyoshi Hamada, Malcolm A.S. Moore

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Figure 1

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(a) Experimental design for “closed system” studies of transmembrane (mi...
(a) Experimental design for “closed system” studies of transmembrane (middle) and transendothelial (right) chemotaxis of CAFCs and LTC-ICs in CD34+ populations placed in the upper chamber of a transwell system, with or without a monolayer of endothelial cells (BMEC-1), with MS-5 cells in the lower chamber as a source of SDF-1 and as a support for CAFCs and LTC-ICs in long-term (5-week) cultures. (b) Experimental design for “open system” studies transmembrane and transendothelial chemotaxis of CAFCs and LTC-ICs in CD34+ populations placed in the upper chamber of a transwell system, with or without a monolayer of endothelial cells (BMEC-1), and with recombinant SDF-1 in the lower chamber (middle). CAFCs and LTC-ICs are assayed by recovery of cells from the upper and lower chambers and secondary transfer to MS-5 monolayers for long-term (5-week) cultures (right). (c) Design as in b but substituting an established monolayer of MS-5 in the lower chamber as a source of SDF-1.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

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Referenced in 23 patents
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