Effective Transduction of Human Adipose Stromal Cells by a Recombinant Adeno-Associated Virus
- Authors: Shevchenko EK1, Makarevich PI2, Tsokolaeva ZI1, Ratner EI1, Parfionova EV3
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Affiliations:
- The Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical Care
- The Faculty of Fundamental Medicine, Lomonosov Moscow State University
- The Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical CareThe Faculty of Fundamental Medicine, Lomonosov Moscow State University
- Issue: Vol 5, No 1 (2010)
- Pages: 60-64
- Section: Articles
- URL: https://genescells.ru/2313-1829/article/view/121496
- DOI: https://doi.org/10.23868/gc121496
- ID: 121496
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Abstract
Human ASC were isolated from adipose tissue obtained from different donors during surgical operations. Low passaged cells were transduced using gene delivery system CStratagene] based on recombinant adeno-associated virus (rAAVJ serotype 2 encoding human vascular endothelial growth factor (VEGF) or green fluorescent protein (GFP). Transduction efficiencies and transgene expression level in ASCs were analyzed by quantitative flow cytometry and ELISA.
ASC population was analysed for heparan sulfate proteoglycan expression, the main cellular AAV binding receptor. It was found that 55-65% of human ASC population express this receptor. The efficiency of ASC transduction using AAV delivery system was found to be 60+7%. GFP expression was visible during a month. Relative to control, cells transduced by VEGF rAAV vector increased VEGF secretion level by at least 20-30 fold as compared to unmanipulated ASC.
Recombinant adeno-associated virus provides efficient tools for ex vivo modification of human ASCs.
Keywords
About the authors
E K Shevchenko
The Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical CareThe Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical Care
P I Makarevich
The Faculty of Fundamental Medicine, Lomonosov Moscow State UniversityThe Faculty of Fundamental Medicine, Lomonosov Moscow State University
Z I Tsokolaeva
The Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical CareThe Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical Care
E I Ratner
The Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical CareThe Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical Care
E V Parfionova
The Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical CareThe Faculty of Fundamental Medicine, Lomonosov Moscow State UniversityThe Institute of Experimental Cardiology of the FSI Russian Cardiologic Research-and-Production Complex of the Federal Agency of High-Tech Medical CareThe Faculty of Fundamental Medicine, Lomonosov Moscow State University
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