In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase
| Autor: | Dan S. Kaufman, R S McIvor, Leonard D. Shultz, Jennifer L. Gori, Roland Gunther, Xinghui Tian, Debra Swanson |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Genetic enhancement
Cellular differentiation Drug Resistance Mice SCID Article methotrexate Cell Line Mice 03 medical and health sciences 0302 clinical medicine dihydrofolate reductase Bone Marrow Mice Inbred NOD In vivo Dihydrofolate reductase Genetics medicine Animals Humans Molecular Biology 030304 developmental biology 0303 health sciences biology Graft Survival Teratoma Cell Differentiation Genetic Therapy embryonic stem cells Molecular biology Embryonic stem cell 3. Good health Tetrahydrofolate Dehydrogenase Cell culture 030220 oncology & carcinogenesis biology.protein Molecular Medicine Methotrexate Stem cell Stem Cell Transplantation medicine.drug |
| Zdroj: | Gene therapy |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/gt.2009.131 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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