A novel competitive repopulation strategy to quantitate engraftment of ex vivo manipulated murine marrow cells in submyeloablated hosts

Brandon K. Wyss, Justin L. Meyers, Anthony L. Sinn, Shanbao Cai, Karen Pollok, W. Goebel

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Objective: Standard competitive repopulation assays have proven valuable in evaluating engraftment potential in ablated hosts, permitting comparisons between various test cell populations. However, no similar method exists to compare engraftment of test cells in submyeloablated hosts, which would be helpful given the applications of reduced-intensity conditioning for hematopoietic gene-replacement therapy and other cellular therapies. Here, we developed a novel assay to quantitate engraftment of hematopoietic stem cells in submyeloablated hosts. Materials and Methods: Engraftment of murine marrow cells transduced with retroviral vectors using two separate protocols was compared to engraftment of fresh untreated competitor cells within low-dose radiation-conditioned hosts using a "three-way" marking system, so that test, competitor, and host cell chimerism could be reliably determined posttransplantation. Results: We demonstrate that the repopulating ability of marrow cells transduced using two distinct protocols was reduced ∼10-fold compared to fresh competitor cells in submyeloablated hosts utilizing the novel "three-way" transplant assay. Conclusions: Murine marrow cells transduced using a clinically applicable protocol acquire an engraftment defect in submyeloablated hosts, similar to cells transduced using a research protocol. We conclude that the submyeloablative competitive repopulation assay described here will be of benefit to comparatively assess the engraftment ability of manipulated hematopoietic stem cells using various culture protocols, such as to test the impact of modifications in transduction protocols needed to attain therapeutic levels of gene-corrected blood cells, or the effect of ex vivo expansion protocols on engraftment potential.

Original languageEnglish
Pages (from-to)513-521
Number of pages9
JournalExperimental Hematology
Volume36
Issue number4
DOIs
StatePublished - Apr 2008

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Bone Marrow
Hematopoietic Stem Cells
Chimerism
Genetic Therapy
Blood Cells
Radiation
Transplants
Therapeutics
Research
Population
Genes

ASJC Scopus subject areas

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

Cite this

A novel competitive repopulation strategy to quantitate engraftment of ex vivo manipulated murine marrow cells in submyeloablated hosts. / Wyss, Brandon K.; Meyers, Justin L.; Sinn, Anthony L.; Cai, Shanbao; Pollok, Karen; Goebel, W.

In: Experimental Hematology, Vol. 36, No. 4, 04.2008, p. 513-521.

Research output: Contribution to journalArticle

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