Highly efficient lentiviral transduction of phenotypically and genotypically characterized endothelial progenitor cells from adult peripheral blood
| Autor: | Boris Fehse, Olga Shardakova, Guenther Giers, Kristoffer Weber, Volker R. Stoldt, Marcus Stockschlaeder, Ruediger E. Scharf |
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| Rok vydání: | 2010 |
| Předmět: |
Adult
Male Genotype Stem Cells Genetic transfer Lentivirus Genetic transduction Endothelial Cells Hematology General Medicine Biology Middle Aged Molecular biology Peripheral blood mononuclear cell Cell biology Gene expression profiling Endothelial stem cell Vasculogenesis Phenotype Transduction Genetic Humans Female Stem cell Progenitor cell Cells Cultured |
| Zdroj: | Blood coagulationfibrinolysis : an international journal in haemostasis and thrombosis. 21(5) |
| ISSN: | 1473-5733 |
| Popis: | Postnatal vasculogenesis has been implicated as an important mechanism for neovascularization via bone marrow-derived endothelial progenitor cells (EPCs) circulating in peripheral blood. In preparation of the utilization of EPCs in clinical protocols, we have generated blood-derived EPCs according to two established protocols by culturing either nonadherent mononuclear cells on fibronectin or adherent mononuclear cells on collagen. To explore the feasibility of these EPCs for their potential clinical use as target cells for genetic transduction to enhance their thromboresistance, newly designed retroviral and lentiviral gene ontology expression vectors were tested. Whereas cell clusters derived from the nonadherent cells demonstrated an only limited proliferative potential, cell colonies derived from collagen-adherent cells expanded more than a million-fold. Characterization of the exponentially growing cells by surface antigen and gene expression profiling revealed a consistently strong expression of characteristic endothelial markers, whereas expression of leukocyte markers was gradually lost. Using a single-step transduction protocol, we were able to achieve gene transfer efficiency of up to 99%. Our results suggest that the generated blood-derived EPC population might be attractive target cells for tissue engineering and gene therapy protocols due to their well defined phenotype, extensive proliferative potential, and efficient genetic transducibility, three important qualities that need to be defined prior to any clinical use. |
| Databáze: | OpenAIRE |
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