A new strategy to generate functional insulin-producing cell lines by somatic gene transfer into pancreatic progenitors.
BACKGROUND
There is increasing interest in developing human cell lines to be used to better understand cell biology, but also for drug screening, toxicology analysis and future cell therapy. In the endocrine pancreatic field, functional human beta cell lines are extremely scarce. On the other hand, rodent insulin producing beta cells have been generated during the past years with great success. Many of such cell lines were produced by using transgenic mice expressing SV40T antigen under the control of the insulin promoter, an approach clearly inadequate in human. Our objective was to develop and validate in rodent an alternative transgenic-like approach, applicable to human tissue, by performing somatic gene transfer into pancreatic progenitors that will develop into beta cells.


RESULTS
In this study, rat embryonic pancreases were transduced with recombinant lentiviral vector expressing the SV40T antigen under the control of the insulin promoter. Transduced tissues were next transplanted under the kidney capsule of immuno-incompetent mice allowing insulinoma development from which beta cell lines were established. Gene expression profile, insulin content and glucose dependent secretion, normalization of glycemia upon transplantation into diabetic mice validated the approach to generate beta cell lines.


CONCLUSIONS
Somatic gene transfer into pancreatic progenitors represents an alternative strategy to generate functional beta cell lines in rodent. Moreover, this approach can be generalized to derive cells lines from various tissues and most importantly from tissues of human origin.
Ravassard
Philippe
P
Biotechnology & Biotherapy group Centre de Recherche Institut du Cerveau et de la Moelle CNRS UMR7225, INSERM UMRS795, Université Pierre et Marie Curie, Paris, France. mail: ravassar@chups.jussieu.fr
Emilie Bricout-Neveu
Hazhouz
Yasmine
Y
Pechberty
Severine
S
Mallet
Jacques
J
Czernichow
Paul
P
Scharfmann
Raphael
R
D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors; D007328:Insulin / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D007340:Insulinoma; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D010179:Pancreas / Q000166:cytology*; D011401:Promoter Regions, Genetic; D051381:Rats; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014161:Transduction, Genetic
D007328:Insulin