A Protocol to Enhance INS1E and MIN6 Functionality-The Use of Theophylline.
In vitro research in the field of type I diabetes is frequently limited by the availability of a functional model for islets of Langerhans. This method shows that by the addition of theophylline to the glucose buffers, mouse insulinoma MIN6 and rat insulinoma INS1E pseudo-islets can serve as a model for islets of Langerhans for in vitro research. The effect of theophylline is dose- and cell line-dependent, resulting in a minimal stimulation index of five followed by a rapid return to baseline insulin secretion by reducing glucose concentrations after a first high glucose stimulation. This protocol solves issues concerning in vitro research for type I diabetes as donors and the availability of primary islets of Langerhans are limited. To avoid the limitations of using human donor material, cell lines represent a valid alternative. Many different β cell lines have been reported, but the lack of reproducible responsiveness to glucose stimulation remains a challenge.
Developmental BioEngineering, MIRA Institute of Biomedical Technology and Technical Medicine, University of Twente, Enschede 7522 NB, The Netherlands. m.grootnibbelink@utwente.nl.
Milou
M
Groot Nibbelink
Developmental BioEngineering, MIRA Institute of Biomedical Technology and Technical Medicine, University of Twente, Enschede 7522 NB, The Netherlands. g.marchioli@tue.nl.
Giulia
G
Marchioli
Department of Complex Tissue Regeneration, MERLN Institute for Technology Inspired Regenerative Medicine, Maastricht University, Maastricht 6200 MD, The Netherlands. l.moroni@maastrichtuniversity.nl.
Lorenzo
L
Moroni
Developmental BioEngineering, MIRA Institute of Biomedical Technology and Technical Medicine, University of Twente, Enschede 7522 NB, The Netherlands. h.b.j.karperien@utwente.nl.
Marcel
M
Karperien
Developmental BioEngineering, MIRA Institute of Biomedical Technology and Technical Medicine, University of Twente, Enschede 7522 NB, The Netherlands. a.a.vanapeldoorn@utwente.nl.
Aart
A
Van Apeldoorn
D000818:Animals; D002460:Cell Line; D003922:Diabetes Mellitus, Type 1 / Q000473:pathology*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008954:Models, Biological; D051381:Rats; D013806:Theophylline / Q000494:pharmacology*
D007328:Insulin; D013806:Theophylline
assay methods; cell lines; insulin secretion in vitro