A Simple High Efficiency Intra-Islet Transduction Protocol Using Lentiviral Vectors

Author(s): Carmen Maria Jimenez-Moreno, Irene de Gracia Herrera-Gomez, Livia Lopez-Noriega, Petra Isabel Lorenzo, Nadia Cobo-Vuilleumier, Esther Fuente-Martin, Jose Manuel Mellado-Gil, Geraldine Parnaud, Domenico Bosco, Benoit Raymond Gauthier, Alejandro Martin-Montalvo.

Journal Name: Current Gene Therapy

Volume 15 , Issue 4 , 2015

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Abstract:

Successful normalization of blood glucose in patients transplanted with pancreatic islets isolated from cadaveric donors established the proof-of-concept that Type 1 Diabetes Mellitus is a curable disease. Nonetheless, major caveats to the widespread use of this cell therapy approach have been the shortage of islets combined with the low viability and functional rates subsequent to transplantation. Gene therapy targeted to enhance survival and performance prior to transplantation could offer a feasible approach to circumvent these issues and sustain a durable functional β-cell mass in vivo. However, efficient and safe delivery of nucleic acids to intact islet remains a challenging task. Here we describe a simple and easy-to-use lentiviral transduction protocol that allows the transduction of approximately 80 % of mouse and human islet cells while preserving islet architecture, metabolic function and glucose-dependent stimulation of insulin secretion. Our protocol will facilitate to fully determine the potential of gene expression modulation of therapeutically promising targets in entire pancreatic islets for xenotransplantation purposes.

Keywords: Diabetes Mellitus, Gene transfer, Infection, Lentivirus, Pancreatic islet, Transduction.

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Article Details

VOLUME: 15
ISSUE: 4
Year: 2015
Page: [436 - 446]
Pages: 11
DOI: 10.2174/1566523215666150630121557

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