Methotrexate supports in vivo selection of human embryonic stem cell derived-hematopoietic cells expressing dihydrofolate reductase

Jennifer L. Gori, R S Mc Ivor, Dan S Kaufman

Research output: Contribution to journalArticle

Abstract

Human embryonic stem cells (hESCs) are an attractive alternative cell source for hematopoietic gene therapy applications as the cells are easily modified with lentiviral or other vectors and can be subsequently induced to differentiate into hematopoietic progenitor cells. However, demonstration of the full hematopoietic potential of hESC-derived progeny is challenging due to low marrow engraftment and the difficulty of detecting cells in the peripheral blood of human/mouse xenografts. Methotrexate (MTX) chemotherapy coupled with expression of a drug resistant dihydrofolate reductase such as Tyr22 (Tyr22DHFR) has the potential to selectively increase engraftment of gene-modified human hematopoietic cells in mice, which would allow for better phenotypic characterization of hESC-derived cells in vivo. We showed that hESCs transduced with Tyr22DHFR-GFP encoding lentivirus vectors differentiate into MTX resistant (MTXr) hemato-endothelial cells. MTX treatment of immunodeficient mice infused with Tyr22DHFR hESC-derived hemato-endothelial cells increased the long-term engraftment of human cells in the bone marrow of MTX-treated mice. In contrast to previous studies, these results indicate that MTX administration has the potential to support in vivo selection that is maintained after cessation of treatment. The MTX/Tyr22DHFR system may therefore be useful for enrichment of gene-modified cell populations in human stem cell and gene therapy applications.

Original languageEnglish (US)
JournalBioengineered Bugs
Volume1
Issue number6
DOIs
StatePublished - Dec 20 2010

Fingerprint

Tetrahydrofolate Dehydrogenase
Stem cells
Methotrexate
Cells
Gene therapy
Endothelial cells
Genetic Therapy
Endothelial Cells
Genes
Lentivirus
Withholding Treatment
Chemotherapy
Cell- and Tissue-Based Therapy
Hematopoietic Stem Cells
Heterografts
Bone Marrow Cells
Human Embryonic Stem Cells
Oxidoreductases
Bone
Blood

Keywords

  • Dihydrofolate reductase
  • Drug resistance
  • Gene therapy
  • Human embryonic stem cells
  • In vivo selection
  • Methotrexate

Cite this

Methotrexate supports in vivo selection of human embryonic stem cell derived-hematopoietic cells expressing dihydrofolate reductase. / Gori, Jennifer L.; Mc Ivor, R S; Kaufman, Dan S.

In: Bioengineered Bugs, Vol. 1, No. 6, 20.12.2010.

Research output: Contribution to journalArticle

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