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Cyclophosphamide/granulocyte colony-stimulating factor causes selective mobilization of bone marrow hematopoietic stem cells into the blood after M phase of the cell cycle
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Cytokine-mobilized peripheral blood hematopoietic stem cells (MPB HSC) are widely used for transplantation in the treatment of malignancies, but the mechanism of HSC mobilization is unclear. Although many HSC in bone marrow (BM) cycle rapidly and expand their numbers in response to cytoreductive agents, such as cyclophosphamide (CY), and cytokines, such as granulocyte colony-stimulating factor (G-CSF), MPB HSC are almost all in the G0 or G1phase of the cell cycle. This has raised the question of whether a subset of noncycling BM HSC is selectively released, or whether cycling BM HSC are mobilized after M phase, but before the next S phase of the cell cycle. To distinguish between these possibilities, mice were treated with one dose of CY followed by daily doses of G-CSF, and dividing cells were marked by administration of bromodeoxyuridine (BrdU) during the interval that BM HSC are expanding. After CY and 4 days of G-CSF, 98.5% of the 2n DNA content long-term repopulating MPB (LT)-HSC stained positively for BrdU, and therefore derived from cells that divided during the treatment interval. Next, LT-HSC from mice previously treated with a single dose of CY, which kills cycling cells, and 3 daily doses of G-CSF, were nearly all killed by a second dose of CY, suggesting that CY/G-CSF causes virtually all LT-HSC to cycle. Analysis of cyclin D2 messenger RNA (mRNA) expression and total RNA content of MPB HSC suggests that these cells are mostly in G1 phase. After CY/G-CSF treatment, virtually all BM LT-HSC enter the cell cycle; some of these HSC then migrate into the blood, specifically after M phase, and are rapidly recruited to particular hematopoietic organs.
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“To drive expression of TERT, we chose a CMV enhancer/β-actin promoter (actin-rtTA+) because this element was previously shown to be active in stem cells23 and in many epithelial tissues, including skin24,25.”
From Conditional telomerase induction causes proliferation of hair follicle stem cells (2005) · cites this paper as [23]
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Metadata
- Authors
- Wright, D. E., Cheshier, S. H., Wagers, A. J., Randall, T. D., Christensen, J. L., & Weissman, I. L.
- Affiliations
- Stanford Medicine (US)
- Venue
- Blood
- Publisher
- American Society of Hematology
- Published
- 2001-04-15
- Volume
- 97
- Issue
- 8
- Pages
- 2278-2285
- ISSN
- 0006-4971 (ISSN-L)
- 1528-0020
- OpenAlex
- W2013051128
- Cited by
- 208
- References
- 58
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 3 · 2023: 7 · 2022: 5 · 2021: 6 · 2020: 4 · 2019: 7 · 2018: 1 · 2017: 4 · 2016: 8 · 2015: 12 · 2014: 7 · 2013: 7 · 2012: 9
- FWCI
- 5.31
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Blood (publishedVersion)
- PubMed (publishedVersion)
- Funders
- American Cancer Society
- Amgen
- National Institutes of Health
- School of Medicine, Stanford University
- National Heart, Lung, and Blood Institute (5R01 HL58770, R01HL058770)
- National Institute of General Medical Sciences (5T32 GM-07365, T32GM007365)
- National Institute of Allergy and Infectious Diseases (5 T32 AI07290, T32AI007290)
- Topics
- Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Mesenchymal stem cell research
- Health Sciences › Medicine › Hematology › Blood disorders and treatments
- Keywords
- hematopoietic stem cell mobilization, cyclophosphamide, granulocyte colony-stimulating factor, bone marrow hematopoietic stem cells, cell cycle, M phase, BrdU labeling, cyclin D2, G1 phase, hematopoietic stem cell transplantation
- MeSH terms
- Animals, Blood Cells, Bone Marrow, Cell Movement, Cells, Cultured, Cyclophosphamide, DNA Replication, Hematopoietic Stem Cells, Metaphase, Mice, Inbred C3H, Mice, Inbred C57BL, Mice, Transgenic, Organ Specificity, Recombinant Fusion Proteins, RNA, Messenger, Spleen, Granulocyte Colony-Stimulating Factor, Cyclins, Cell Lineage, Hematopoietic Stem Cell Mobilization, Mice, Cyclin D2
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