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Transduction-Transplantation Mouse Model of Myeloproliferative Neoplasm
Published on: December 22, 2016
Optimized transduction of canine paediatric CD34(+) cells using an MSCV-based bicistronic vector.
S E Suter1, T A Gouthro, P A McSweeney
1Section of Medical Genetics, Department of Clinical Studies, Philadelphia, Pennsylvania, USA. sesuter@vmth.ucdavis.edu
Veterinary Research Communications
|December 2, 2006
Summary
This study optimized retroviral transduction in canine cells, achieving high efficiency in fibroblasts and CD34(+) bone marrow cells for potential genetic disease treatments.
Area of Science:
- * Gene therapy and retroviral vector development.
- * Canine molecular biology and immunology.
Background:
- * Developing efficient gene delivery methods is crucial for treating genetic diseases in canines.
- * Canine X-linked severe combined immunodeficiency requires effective gene therapy strategies.
Purpose of the Study:
- * To optimize retroviral transduction of canine cells using a bicistronic vector.
- * To evaluate transduction efficiency in canine fibroblasts and CD34(+) bone marrow cells.
Main Methods:
- * Utilized a murine MSCV-based bicistronic retroviral vector encoding canine common gamma chain (gammac) and enhanced green fluorescent protein (EGFP).
- * Employed amphotropic virus producer cell lines (PA317 and Phoenix) for virus production.
- * Optimized transduction using centrifugation (spinfection) with polybrene and Retronectin-coated plates.
Main Results:
- * Achieved approximately 75% transduction efficiency in Cf2Th canine fibroblast cells.
- * Transduced an average of 11% of canine CD34(+) bone marrow cells using spinfection and Retronectin.
- * Confirmed expression of functional canine gammac protein on the surface of transduced cells.
Conclusions:
- * The optimized retroviral transduction protocol is effective for canine cells, including primary CD34(+) cells.
- * This method shows promise for treating canine genetic disorders like X-linked severe combined immunodeficiency.
- * Successful transduction of sufficient CD34(+) cells could enable gene therapy for various canine diseases.
