title: A Simple High Efficiency Intra-Islet Transduction Protocol Using Lentiviral Vectors.
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.
authors.forename: Carmen Maria
authors.initials: CM
authors.lastname: Jimenez-Moreno
authors.forename: Irene de Gracia
authors.initials: IG
authors.lastname: Herrera-Gomez
authors.forename: Livia
authors.initials: L
authors.lastname: Lopez-Noriega
authors.forename: Petra Isabel
authors.initials: PI
authors.lastname: Lorenzo
authors.forename: Nadia
authors.initials: N
authors.lastname: Cobo-Vuilleumier
authors.forename: Esther
authors.initials: E
authors.lastname: Fuente-Martin
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Mellado-Gil
authors.forename: Geraldine
authors.initials: G
authors.lastname: Parnaud
authors.forename: Domenico
authors.initials: D
authors.lastname: Bosco
authors.affiliation: Pancreatic Islet Development and Regeneration Unit, Department of Stem Cells, CABIMER-Andalusian Center for Molecular Biology and Regenerative Medicine, Avenida Americo Vespucio, Parque Científico y Tecnologico Cartuja 93, 41092 Sevilla, Spain. benoit.gauthier@cabimer.es.
authors.forename: Benoit Raymond
authors.initials: BR
authors.lastname: Gauthier
authors.forename: Alejandro
authors.initials: A
authors.lastname: Martin-Montalvo
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005434:Flow Cytometry; D005822:Genetic Vectors; D005934:Glucagon / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000502:physiology*; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D008810:Mice, Inbred C57BL; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D007328:Insulin; D049452:Green Fluorescent Proteins; D005934:Glucagon