Efficient gene delivery and silencing of mouse and human pancreatic islets

BMC Biotechnology

Methodology Article

Kathrin Maedler¹  Bruno Lukowiak²  Julie Kerr-conte²  Justine Longue²  Bruno Lefebvre²  Brigitte Vandewalle²  Ericka Moerman²  Valery Gmyr²  François Pattou³ 

[1] Center for Biomolecular Interactions Bremen, University of Bremen, Germany;Univ Lille Nord de France, F-59000, Lille, France;INSERM U859, F-59000, Lille, France;Univ Lille Nord de France, F-59000, Lille, France;INSERM U859, F-59000, Lille, France;CHU Lille, F-59000, Lille, France;

关键词: Beta Cell;    Human Islet;    Insulin Content;    Dithizone;    Mouse Islet;   

DOI  :  10.1186/1472-6750-10-28

 received in 2009-10-20, accepted in 2010-03-30,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundIn view of the importance of beta cells in glucose homeostasis and the profound repercussions of beta cell pathology on human health, the acquisition of tools to study pancreatic islet function is essential for the design of alternative novel therapies for diabetes. One promising approach toward this goal involves the modification of gene expression profile of beta cells.ResultsThis study describes a new method of gene and siRNA delivery into human pancreatic islets by microporation technology. We demonstrated that mild islet distention with accutase greatly enhanced the transfection efficiency without compromising in vitro function (secretion, apoptosis and viability). As an example, the recently identified gene involved in type 2 diabetes, ZnT8, can be over-expressed or silenced by RNA interference using this technology. Microporation can also be used on rodent islets.ConclusionsTaken together, our results demonstrate that microporation technology can be used to modify gene expression in whole rodent and human islets without altering their in vitro function and will be key to the elucidation of the factors responsible for proper islet function.

【 授权许可】

CC BY   
© Lefebvre et al; licensee BioMed Central Ltd. 2010