Homing of purified murine lymphohematopoietic stem cells: A cytokine-induced defect

Jan Cerny, Mark Dooner, Christina McAuliffe, Houri Habibian, Kimberly Stencil, Virla Berrios, Judy Reilly, Jane Carlson, Anna Maria Cerny, Lionel D'Hondt, Brian Benoit, Jean Francois Lambert, Gerald Colvin, Susan Nilsson, Pamela Becker, Peter Quesenberry

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Abstract

This study was designed to establish a direct homing assay using purified lineage-negative Sca-1-positive (Lin-Sca+) murine bone marrow cells and to evaluate the effects of cytokines on homing. C57BL/6 Lin-Sca+ marrow stem cells were labeled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and then injected by tail vein into untreated C57BL/6 mice. Marrow was harvested at various times after cell infusion and analyzed on a high-speed MoFlo cell sorter for fluorescent positive events, using a large event analysis, with at least 16 million total events analyzed. We have shown that homing of Lin- Sca+ cells plateaus by I h, and at 3 h post-infusion is linear between 50,000 and 1,000,000 infused cells. This forms a base for a homing assay in which 250,000 CFDA-SE labeled Lin- Sca+ marrow cells are infused and then recovered from marrow 3 h later, followed by a large-event fluorescence-activated cell sorting (FACS) analysis. We found that 7.45-9.32% of infused cells homed and that homing of stem cells cultured for 48 h in interleukin-3 (IL-3), IL-6, IL-11, and steel factor cultured cells was defective when compared to noncultured cells. Exposure of marrow stem cells to IL-3, IL-6, IL-11, and steel factor induces a stem cell homing defect, which probably underlies the engraftment defect previously characterized under these conditions.

Original languageEnglish
Pages (from-to)913-922
Number of pages10
JournalJournal of Hematotherapy and Stem Cell Research
Volume11
Issue number6
DOIs
Publication statusPublished - Dec 2002
Externally publishedYes

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