Marrow repopulating ability of SCA-I+ LIN- cells fractionated on the basis of adhesion molecule expression and position in cell cycle

C. M. Travcoff, R. Dagher, Mervin Yoder, K. Hiatt

Research output: Contribution to journalArticle

Abstract

Sea-1 lin cells capable of providing long-term engraftment in lethally irradiated recipients can be fractionated based on their high expression of CD43, CD44, and CD49e, and low or lack of expression of CD1 la, CD49d, and CD62L. In vitro studies demonstrate that these populations of Sca-1 in- engrafting ceils possess the majority of HPP-CFC, LPPCFC, and cells providing rapid in vitro expansion. However, based on the belief that primitive hematopoietic progenitor cells (HPC) represent a relatively quiescent population of stem cells, Sca-l+ lin" CD43+, CD44+, or CD49e+ (adhesion) cells would not be expected to be enriched for engrafting cells, since these three phenotypes contain a relatively high percentage of cycling cells (30% to 50% in S/G2+M) compared to their corresponding adhesion molecule-negative counterparts. This posed the question of whether primitive HPC were predominantly contained within the quiescent (Ga/G \) or mitotically active (S/G2+M) fractions of Sea-1+ lin" adhesion+ cells. To begin to investigate this, the three groups of Sca-l+ lin" adhésion cells were individually isolated and further fractionated based on their position in the cell cycle with the DNA dye Hoechst 33342. In vitro analysis revealed marked proliferative potential of both quiescent and cycling Sca-l+ lin" adhésion cells. To evaluate engraftment potential, a total of 103 Sca-l+ lin- adhésion Grj/Gi or S/G2+M cells isolated from SJL mice were transplanted into lethally irradiated C57/BI6 recipients along with 3 x 104 C57B1/6 competitor cells. The percentage of CD45.2 peripheral blood cells was used to estimate chimerism. Analysis of engraftment up to 3 months posttransplantation revealed the ability of cycling Sea-1+lin" adhésion cells to provide both myelopoiesis and lymphopoiesis in transplanted recipients. These data suggest that expression of CD43, CD44, and CD49e, independent of cell cycle status, may be a useful predictor of the engraftment potential of murine hematopoietic stem cells.

Original languageEnglish
Pages (from-to)697
Number of pages1
JournalExperimental Hematology
Volume26
Issue number8
StatePublished - 1998

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Cell Cycle
Bone Marrow
Hematopoietic Stem Cells
Oceans and Seas
Cell Adhesion
Lymphopoiesis
Myelopoiesis
Chimerism
Population
Blood Cells
Coloring Agents
Stem Cells
Phenotype
DNA
In Vitro Techniques

ASJC Scopus subject areas

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

Cite this

Marrow repopulating ability of SCA-I+ LIN- cells fractionated on the basis of adhesion molecule expression and position in cell cycle. / Travcoff, C. M.; Dagher, R.; Yoder, Mervin; Hiatt, K.

In: Experimental Hematology, Vol. 26, No. 8, 1998, p. 697.

Research output: Contribution to journalArticle

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abstract = "Sea-1 lin cells capable of providing long-term engraftment in lethally irradiated recipients can be fractionated based on their high expression of CD43, CD44, and CD49e, and low or lack of expression of CD1 la, CD49d, and CD62L. In vitro studies demonstrate that these populations of Sca-1 in- engrafting ceils possess the majority of HPP-CFC, LPPCFC, and cells providing rapid in vitro expansion. However, based on the belief that primitive hematopoietic progenitor cells (HPC) represent a relatively quiescent population of stem cells, Sca-l+ lin{"} CD43+, CD44+, or CD49e+ (adhesion) cells would not be expected to be enriched for engrafting cells, since these three phenotypes contain a relatively high percentage of cycling cells (30{\%} to 50{\%} in S/G2+M) compared to their corresponding adhesion molecule-negative counterparts. This posed the question of whether primitive HPC were predominantly contained within the quiescent (Ga/G \) or mitotically active (S/G2+M) fractions of Sea-1+ lin{"} adhesion+ cells. To begin to investigate this, the three groups of Sca-l+ lin{"} adh{\'e}sion cells were individually isolated and further fractionated based on their position in the cell cycle with the DNA dye Hoechst 33342. In vitro analysis revealed marked proliferative potential of both quiescent and cycling Sca-l+ lin{"} adh{\'e}sion cells. To evaluate engraftment potential, a total of 103 Sca-l+ lin- adh{\'e}sion Grj/Gi or S/G2+M cells isolated from SJL mice were transplanted into lethally irradiated C57/BI6 recipients along with 3 x 104 C57B1/6 competitor cells. The percentage of CD45.2 peripheral blood cells was used to estimate chimerism. Analysis of engraftment up to 3 months posttransplantation revealed the ability of cycling Sea-1+lin{"} adh{\'e}sion cells to provide both myelopoiesis and lymphopoiesis in transplanted recipients. These data suggest that expression of CD43, CD44, and CD49e, independent of cell cycle status, may be a useful predictor of the engraftment potential of murine hematopoietic stem cells.",
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T1 - Marrow repopulating ability of SCA-I+ LIN- cells fractionated on the basis of adhesion molecule expression and position in cell cycle

AU - Travcoff, C. M.

AU - Dagher, R.

AU - Yoder, Mervin

AU - Hiatt, K.

PY - 1998

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N2 - Sea-1 lin cells capable of providing long-term engraftment in lethally irradiated recipients can be fractionated based on their high expression of CD43, CD44, and CD49e, and low or lack of expression of CD1 la, CD49d, and CD62L. In vitro studies demonstrate that these populations of Sca-1 in- engrafting ceils possess the majority of HPP-CFC, LPPCFC, and cells providing rapid in vitro expansion. However, based on the belief that primitive hematopoietic progenitor cells (HPC) represent a relatively quiescent population of stem cells, Sca-l+ lin" CD43+, CD44+, or CD49e+ (adhesion) cells would not be expected to be enriched for engrafting cells, since these three phenotypes contain a relatively high percentage of cycling cells (30% to 50% in S/G2+M) compared to their corresponding adhesion molecule-negative counterparts. This posed the question of whether primitive HPC were predominantly contained within the quiescent (Ga/G \) or mitotically active (S/G2+M) fractions of Sea-1+ lin" adhesion+ cells. To begin to investigate this, the three groups of Sca-l+ lin" adhésion cells were individually isolated and further fractionated based on their position in the cell cycle with the DNA dye Hoechst 33342. In vitro analysis revealed marked proliferative potential of both quiescent and cycling Sca-l+ lin" adhésion cells. To evaluate engraftment potential, a total of 103 Sca-l+ lin- adhésion Grj/Gi or S/G2+M cells isolated from SJL mice were transplanted into lethally irradiated C57/BI6 recipients along with 3 x 104 C57B1/6 competitor cells. The percentage of CD45.2 peripheral blood cells was used to estimate chimerism. Analysis of engraftment up to 3 months posttransplantation revealed the ability of cycling Sea-1+lin" adhésion cells to provide both myelopoiesis and lymphopoiesis in transplanted recipients. These data suggest that expression of CD43, CD44, and CD49e, independent of cell cycle status, may be a useful predictor of the engraftment potential of murine hematopoietic stem cells.

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