In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase.
Document Type
Article
Publication Date
2010
Keywords
Bone-Marrow, Cell-Differentiation, Cell-Line, Drug-Resistance, Embryonic-Stem-Cells, Gene-Therapy, Graft-Survival, Humans, Methotrexate, Mice-Inbred-NOD, Mice-SCID, Stem-Cell-Transplantation, Teratoma, Tetrahydrofolate-Dehydrogenase
First Page
238
Last Page
249
JAX Location
see Reprint Collection (a pdf is available).
JAX Source
Gene Ther 2010 Feb; 17(2):238-49.
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-2Rgammac(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.
Recommended Citation
Gori JL,
Tian X,
Swanson D,
Gunther R,
Shultz LD,
McIvor RS,
Kaufman DS.
In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase. Gene Ther 2010 Feb; 17(2):238-49.