We studied the genetic and engraftment phenotype of highly purified murine hematopoietic stem cells (lineage negative, rhodamine-low, Hoechst-low) through cytokine-stimulated cell cycle. Cells were cultured in interleukin (IL)-3, IL-6, IL-11, and steel factor for 0 to 48 h and tested for engraftment capacity in a lethally irradiated murine competitive transplant model. Engraftment showed major fluctuations with nadirs at 36 and 48 h of culture and recovery during the next G1. Gene expression of quiescent (0 h) or cycling (48 h) stem cells was compared with lineage positive cells by 3′ end PCR differential display analysis. Individual PCR bands were quantified using a 0 to 9 scale and results were visually compared using color-coded matrices. We defined a set of 637 transcripts expressed in stem cells and not expressed in lineage positive cells. Gene expression analyzed at 0 and 48 h showed a major shift from “stem cell genes” being highly expressed at 0 h and turned off at 48 h, while “cell division” genes were turned on at 48 h. These observations suggest stem cell gene expression shifts through cell cycle in relation to cell cycle related alterations of stem cell phenotype. The engraftment defect is related to a major phenotypic change of the stem cell.
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2 June 2003
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June 02 2003
Marrow Stem Cells Shift Gene Expression and Engraftment Phenotype with Cell Cycle Transit
Jean-François Lambert,
Jean-François Lambert
1Roger Williams Medical Center, Providence, RI 02908
2Division of Hematology, University Hospital, CH1211 Geneva 14, Switzerland
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Meng Liu,
Meng Liu
3Schepens Eye Research Institute, Harvard University, Boston, MA 02114
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Gerald A. Colvin,
Gerald A. Colvin
1Roger Williams Medical Center, Providence, RI 02908
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Mark Dooner,
Mark Dooner
1Roger Williams Medical Center, Providence, RI 02908
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Christina I. McAuliffe,
Christina I. McAuliffe
1Roger Williams Medical Center, Providence, RI 02908
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Pamela S. Becker,
Pamela S. Becker
5University of Massachusetts Cancer Center, Worcester, MA 01655
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Bernard G. Forget,
Bernard G. Forget
4Department of Genetics, Yale University School of Medicine, New Haven, CT 06520
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Sherman M. Weissman,
Sherman M. Weissman
4Department of Genetics, Yale University School of Medicine, New Haven, CT 06520
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Peter J. Quesenberry
Peter J. Quesenberry
1Roger Williams Medical Center, Providence, RI 02908
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Jean-François Lambert
1Roger Williams Medical Center, Providence, RI 02908
2Division of Hematology, University Hospital, CH1211 Geneva 14, Switzerland
Meng Liu
3Schepens Eye Research Institute, Harvard University, Boston, MA 02114
Gerald A. Colvin
1Roger Williams Medical Center, Providence, RI 02908
Mark Dooner
1Roger Williams Medical Center, Providence, RI 02908
Christina I. McAuliffe
1Roger Williams Medical Center, Providence, RI 02908
Pamela S. Becker
5University of Massachusetts Cancer Center, Worcester, MA 01655
Bernard G. Forget
4Department of Genetics, Yale University School of Medicine, New Haven, CT 06520
Sherman M. Weissman
4Department of Genetics, Yale University School of Medicine, New Haven, CT 06520
Peter J. Quesenberry
1Roger Williams Medical Center, Providence, RI 02908
Address correspondence to Peter J. Quesenberry, Director, Department of Research, Roger Williams Medical Center, 825 Chalkstone Ave., Providence, RI 02908. Phone: 401-456-5770; Fax: 401-456-5759; E-mail: [email protected]
*
Abbreviation used in this paper: SCF, stem cell factor (or steel factor).
Received:
January 09 2003
Revision Received:
April 08 2003
Accepted:
April 16 2003
Online ISSN: 1540-9538
Print ISSN: 0022-1007
The Rockefeller University Press
2003
J Exp Med (2003) 197 (11): 1563–1572.
Article history
Received:
January 09 2003
Revision Received:
April 08 2003
Accepted:
April 16 2003
Citation
Jean-François Lambert, Meng Liu, Gerald A. Colvin, Mark Dooner, Christina I. McAuliffe, Pamela S. Becker, Bernard G. Forget, Sherman M. Weissman, Peter J. Quesenberry; Marrow Stem Cells Shift Gene Expression and Engraftment Phenotype with Cell Cycle Transit . J Exp Med 2 June 2003; 197 (11): 1563–1572. doi: https://doi.org/10.1084/jem.20030031
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