Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells

C. L. Miller, V. I. Rebel, M. E. Lemieux, C. D. Helgason, P. M. Lansdorp, C. J. Eaves

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Previous studies have suggested that Steel factor (SF) can influence the behavior of many types of hematopoietic progenitor cells both in vivo and in vitro, although whether these may include the most primitive populations of totipotent repopulating cells remains controversial. To approach this question, we measured the number of Sca1+Lin-WGA+ cells, the number of cells with demonstrable myeloid (long-term culture-initiating cell [LTC-IC]) or both myeloid and lymphoid (LTC-IC(ML)) potential in 4- to 5-week-old long-term cultures containing irradiated primary marrow feeder layers, and the number of multilineage long-term in vivo repopulating cells (competitive repopulating unit [CRU]) present in the marrow of W42/+ or W41/W41 mice compared to +/+ controls. There was no significant effect of either of these W mutations on the number of Sca1+Lin-WGA+ cells and, in W41/W41 mice, neither LTC-IC nor LTC-IC(ML) populations appeared to be affected. On the other hand, although W41/W41 and W42/+ cells could both be detected in the in vivo CRU assay, their numbers were markedly reduced (17- and seven-fold, respectively) in spite of the fact that both of these W mutant genotypes contained near normal numbers of day-9 and -12 colony-forming units-spleen (CFU-S). In vitro quantitation of erythroid (burst-forming units-erythroid [BFU-E]), granulopoietic (CFU-granulocyte/macrophage [CFU-GM]), multilineage (CFU-granulocyte/erythrocyte/monocyte/macrophage [CFU-GEMM]), and pre-B clonogenic progenitors (CFU-pre-B) also revealed no differences in the numbers (or proliferative potential) of any of these cells when W41/W41 or W42/+ and normal mice were compared, although day 3 BFU-E from both types of W mutant mice showed no response to the typical enhancing effect exerted by SF on their +/+ counterparts. Taken together, these findings are consistent with the view that SF activation of c-kit receptor-induced signaling events is not a rate-limiting mechanism controlling red blood cell production during normal development until hematopoietic cells differentiate beyond the day-3 BFU-E stage. Nevertheless, normal hematopoietic stem cells do appear to be responsive to SF, since their W mutant counterparts display a disadvantage in the in vivo setting which is exaggerated under conditions of hematopoietic regeneration. On the other hand, alternative mechanisms also appear to contribute to the regulation of hematopoietic stem cell numbers in vivo and to their detection as LTC-IC in vitro.

Original languageEnglish (US)
Pages (from-to)185-194
Number of pages10
JournalExperimental Hematology
Volume24
Issue number2
StatePublished - 1996
Externally publishedYes

Fingerprint

Hematopoietic Stem Cells
Stem Cell Factor
Cell Culture Techniques
Erythroid Precursor Cells
Granulocytes
Cell Count
Erythrocytes
Bone Marrow
Macrophages
Proto-Oncogene Proteins c-kit
Feeder Cells
Population
Regeneration
Monocytes
Stem Cells
Spleen
Genotype
Mutation
In Vitro Techniques

Keywords

  • competitive repopulating unit
  • LTC-IC
  • LTC-IC(ML)
  • Steel factor
  • W/W and W/+ mice

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Miller, C. L., Rebel, V. I., Lemieux, M. E., Helgason, C. D., Lansdorp, P. M., & Eaves, C. J. (1996). Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells. Experimental Hematology, 24(2), 185-194.

Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells. / Miller, C. L.; Rebel, V. I.; Lemieux, M. E.; Helgason, C. D.; Lansdorp, P. M.; Eaves, C. J.

In: Experimental Hematology, Vol. 24, No. 2, 1996, p. 185-194.

Research output: Contribution to journalArticle

Miller, CL, Rebel, VI, Lemieux, ME, Helgason, CD, Lansdorp, PM & Eaves, CJ 1996, 'Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells', Experimental Hematology, vol. 24, no. 2, pp. 185-194.
Miller CL, Rebel VI, Lemieux ME, Helgason CD, Lansdorp PM, Eaves CJ. Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells. Experimental Hematology. 1996;24(2):185-194.
Miller, C. L. ; Rebel, V. I. ; Lemieux, M. E. ; Helgason, C. D. ; Lansdorp, P. M. ; Eaves, C. J. / Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells. In: Experimental Hematology. 1996 ; Vol. 24, No. 2. pp. 185-194.
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T1 - Studies of W mutant mice provide evidence for alternate mechanisms capable of activating hematopoietic stem cells

AU - Miller, C. L.

AU - Rebel, V. I.

AU - Lemieux, M. E.

AU - Helgason, C. D.

AU - Lansdorp, P. M.

AU - Eaves, C. J.

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N2 - Previous studies have suggested that Steel factor (SF) can influence the behavior of many types of hematopoietic progenitor cells both in vivo and in vitro, although whether these may include the most primitive populations of totipotent repopulating cells remains controversial. To approach this question, we measured the number of Sca1+Lin-WGA+ cells, the number of cells with demonstrable myeloid (long-term culture-initiating cell [LTC-IC]) or both myeloid and lymphoid (LTC-IC(ML)) potential in 4- to 5-week-old long-term cultures containing irradiated primary marrow feeder layers, and the number of multilineage long-term in vivo repopulating cells (competitive repopulating unit [CRU]) present in the marrow of W42/+ or W41/W41 mice compared to +/+ controls. There was no significant effect of either of these W mutations on the number of Sca1+Lin-WGA+ cells and, in W41/W41 mice, neither LTC-IC nor LTC-IC(ML) populations appeared to be affected. On the other hand, although W41/W41 and W42/+ cells could both be detected in the in vivo CRU assay, their numbers were markedly reduced (17- and seven-fold, respectively) in spite of the fact that both of these W mutant genotypes contained near normal numbers of day-9 and -12 colony-forming units-spleen (CFU-S). In vitro quantitation of erythroid (burst-forming units-erythroid [BFU-E]), granulopoietic (CFU-granulocyte/macrophage [CFU-GM]), multilineage (CFU-granulocyte/erythrocyte/monocyte/macrophage [CFU-GEMM]), and pre-B clonogenic progenitors (CFU-pre-B) also revealed no differences in the numbers (or proliferative potential) of any of these cells when W41/W41 or W42/+ and normal mice were compared, although day 3 BFU-E from both types of W mutant mice showed no response to the typical enhancing effect exerted by SF on their +/+ counterparts. Taken together, these findings are consistent with the view that SF activation of c-kit receptor-induced signaling events is not a rate-limiting mechanism controlling red blood cell production during normal development until hematopoietic cells differentiate beyond the day-3 BFU-E stage. Nevertheless, normal hematopoietic stem cells do appear to be responsive to SF, since their W mutant counterparts display a disadvantage in the in vivo setting which is exaggerated under conditions of hematopoietic regeneration. On the other hand, alternative mechanisms also appear to contribute to the regulation of hematopoietic stem cell numbers in vivo and to their detection as LTC-IC in vitro.

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