In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase

J. L. Gori, X. Tian, D. Swanson, R. Gunther, L. D. Shultz, R S Mc Ivor, Dan S Kaufman

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Human embryonic stem cells (hESCs) provide a novel source of hematopoietic and other cell populations suitable for gene therapy applications. Preclinical studies to evaluate engraftment of hESC-derived hematopoietic cells transplanted into immunodeficient mice demonstrate only limited repopulation. Expression of a drug-resistance gene, such as Tyr22-dihydrofolate reductase (Tyr22-DHFR), coupled to methotrexate (MTX) chemotherapy has the potential to selectively increase the engraftment of gene-modified, hESC-derived cells in mouse xenografts. Here, we describe the generation of Tyr22-DHFR-GFP-expressing hESCs that maintain pluripotency, produce teratomas and can differentiate into MTXr-hemato-endothelial cells. We demonstrate that MTX administered to nonobese diabetic/severe combined immunodeficient/IL-2Rγc null (NSG) mice after injection of Tyr22-DHFR-hESC-derived cells significantly increases human CD34 and CD45 cell engraftment in the bone marrow (BM) and peripheral blood of transplanted MTX-treated mice. These results demonstrate that MTX treatment supports selective, long-term engraftment of Tyr22-DHFR cells in vivo, and provides a novel approach for combined human cell and gene therapy.

Original languageEnglish (US)
Pages (from-to)238-249
Number of pages12
JournalGene therapy
Volume17
Issue number2
DOIs
StatePublished - Feb 1 2010

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Tetrahydrofolate Dehydrogenase
Methotrexate
Genetic Therapy
Teratoma
Cell- and Tissue-Based Therapy
Drug Resistance
Heterografts
Genes
Human Embryonic Stem Cells
Endothelial Cells
Bone Marrow
Drug Therapy
Injections
Population

Keywords

  • Dihydrofolate reductase
  • Embryonic stem cells
  • Methotrexate

Cite this

In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase. / Gori, J. L.; Tian, X.; Swanson, D.; Gunther, R.; Shultz, L. D.; Mc Ivor, R S; Kaufman, Dan S.

In: Gene therapy, Vol. 17, No. 2, 01.02.2010, p. 238-249.

Research output: Contribution to journalArticle

Gori, J. L. ; Tian, X. ; Swanson, D. ; Gunther, R. ; Shultz, L. D. ; Mc Ivor, R S ; Kaufman, Dan S. / In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase. In: Gene therapy. 2010 ; Vol. 17, No. 2. pp. 238-249.
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