A Simple High Efficiency Intra-Islet Transduction Protocol Using Lentiviral Vectors
Carmen Maria Jimenez-Moreno,
Irene de Gracia Herrera-Gomez,
Petra Isabel Lorenzo,
Jose Manuel Mellado-Gil,
Benoit Raymond Gauthier,
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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