Marrow stem cells shift gene expression and engraftment phenotype with cell cycle transit.
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.
Lambert
Jean-Francois
JF
Department of Research, Roger Williams Medical Center, 825 Chalkstone Ave., Providence, RI 02908, USA.
Liu
Meng
M
Colvin
Gerald A
GA
Dooner
Mark
M
McAuliffe
Christina I
CI
Becker
Pamela S
PS
Forget
Bernard G
BG
Weissman
Sherman M
SM
Quesenberry
Peter J
PJ
D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D017370:Interleukin-11 / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D019089:Stem Cell Factor / Q000494:pharmacology
D018076:DNA, Complementary; D017370:Interleukin-11; D007377:Interleukin-3; D015850:Interleukin-6; D019089:Stem Cell Factor