title: Encapsulation of Insulin-Secreting Cells Expressing a Genetically Encoded Fluorescent Calcium Indicator for Cell-Based Sensing In Vivo.
abstract: The development of cell-based biosensors that give insight into cell and tissue function in vivo is an attractive technology for biomedical research. Here, the development of a cell line expressing a fluorescent calcium sensor for the study of beta-cell function in vivo is reported. The bioresponsive cell model is based on INS-1E pancreatic beta-cells, stably expressing the genetically encoded cameleon-based fluorescent sensor YC3.6cyto. Following single-cell selection and expansion, functional testing and in vitro encapsulation experiments are used to identify a suitable clone of INS-1E cells expressing the calcium sensor. This clone is transplanted subcutaneous in mouse using a cell macroencapsulation system based on flat sheet porous membranes. Cells in the implanted capsules are able to respond to glucose in vivo by secreting insulin and thereby contributing to the regulation of glycaemia in the mice. Furthermore, fluorescence imaging of explanted devices shows that encapsulated cells maintain high level expression of YC3.6cytoin vivo. In conclusion, these data show that encapsulated INS-1E cells stably expressing a genetically encoded calcium sensor can be successfully implanted in vivo, and therefore serve as biosensing element or in vivo model to longitudinally monitor the function of pancreatic beta-cells.
authors.affiliation: Device Engineering, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Christophe
authors.initials: C
authors.lastname: Boss
authors.affiliation: Mitochondrial Function, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Umberto
authors.initials: U
authors.lastname: De Marchi
authors.affiliation: Mitochondrial Function, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Hermant
authors.affiliation: Device Engineering, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Mouna
authors.initials: M
authors.lastname: Conrad
authors.affiliation: Flow Cytometry, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Federico
authors.initials: F
authors.lastname: Sizzano
authors.affiliation: Flow Cytometry, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Alessio
authors.initials: A
authors.lastname: Palini
authors.affiliation: Mitochondrial Function, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Wiederkehr
authors.affiliation: Device Engineering, Nestlé Institute of Health Sciences, EPFL Innovation Park, 1015, Lausanne, Switzerland.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bouche
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D045744:Cell Line, Tumor; D018914:Cells, Immobilized / Q000378:metabolism / Q000637:transplantation; D064593:Heterografts; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism / Q000637:transplantation; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D016513:Mice, SCID; D051381:Rats
chemical_list: D002135:Calcium-Binding Proteins; D007328:Insulin; D008164:Luminescent Proteins; D002118:Calcium
keywords: cell encapsulation; cell-based biosensor; genetically encoded calcium indicator; in vivo fluorescence imaging; pancreatic beta-cells