Transduction of non-dividing adult human pancreatic beta cells by an integrating lentiviral vector.
Pancreatic islet cells are terminally differentiated endocrine cells and are refractory to stable infection by retroviral vectors, which require the breakdown of the nuclear membrane during cell division in order to insert the transgene into the host cell genome. Thus, attempts to render beta-cell allografts less immunogenic have had to rely on stable transfection of surrogate cells. Similarly, this problem has precluded the development of conditionally immortalized human beta cells for clinical allotransplantation. In this report, we demonstrate that adult human islet beta cells can be transduced by a new three-plasmid integrating lentiviral vector with an efficiency of 62 +/- 1.8 % at a multiplicity of infection (MOI) of 2.5 in vitro. This work makes genetic engineering of adult human pancreatic beta cells possible for the first time, allowing strategies to render beta-cell allografts non-immunogenic to be optimized and to creating conditionally immortalized human beta cells for clinical transplantation.
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Diabetes Research Center, Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Edelstein
D
D
Brendel
M D
MD
Brandhorst
D
D
Brandhorst
H
H
Bretzel
R G
RG
Brownlee
M
M
D002455:Cell Division; D002461:Cell Line, Transformed; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism* / Q000821:virology*; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics; D008242:Lysogeny; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012190:Retroviridae / Q000235:genetics; D014162:Transfection / Q000235:genetics
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins