title: Innovative Microcapsules for Pancreatic β-Cells Harvested from Mature Double-Transgenic Mice: Cell Imaging, Viability, Induced Glucose-Stimulated Insulin Measurements and Proinflammatory Cytokines Analysis.
abstract: Recently we demonstrated that microencapsulation of a murine pancreatic β-cell line using an alginate-ursodeoxycholic acid (UDCA) matrix produced microcapsules with good stability and cell viability. In this study, we investigated if translation of this formulation to microencapsulation of primary β-cells harvested from mature double-transgenic healthy mice would also generate stable microcapsules with good cell viability.
  
  
  
  Islets of Langerhans were isolated from Ngn3-GFP/RIP-DsRED mice by intraductal collagenase P digestion and density gradient centrifugation, dissociated into single cells and the β-cell population purified by Fluorescence Activated Cell Sorting. β-cells were microencapsulated using either alginate-poly-l-ornithine (F1; control) or alginate-poly-l-ornithine-UDCA (F2; test) formulations. Microcapsules were microscopically examined and microencapsulated cells were analyzed for viability, insulin and cytokine release, 2 days post-microencapsulation.
  
  
  
  Microcapsules showed good uniformity and morphological characteristics and even cell distribution within microcapsules with or without UDCA. Two days post microencapsulation cell viability, mitochondrial ATP and insulin production were shown to be optimized in the presence of UDCA whilst production of the proinflammatory cytokine IL-1β was reduced. Contradictory to our previous studies, UDCA did not reduce production of any other pro-inflammatory biomarkers.
  
  
  
  These results suggest that UDCA incorporation improves microcapsules' physical and morphological characteristics and improves the viability and function of encapsulated mature primary pancreatic β-cells.
authors.affiliation: Biotechnology and Drug Development Research Laboratory, School of Pharmacy, Curtin Health Innovation Research Institute (CHIRI), Curtin University, Perth, WA, Australia.
authors.forename: Armin
authors.initials: A
authors.lastname: Mooranian
authors.affiliation: School of Public Health, Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia.
authors.forename: Ryu
authors.initials: R
authors.lastname: Tackechi
authors.affiliation: Harry Perkins Institute of Medical Research, Centre for Diabetes Research, Perth, WA, Australia.
authors.forename: Emma
authors.initials: E
authors.lastname: Jamieson
authors.affiliation: Harry Perkins Institute of Medical Research, Centre for Diabetes Research, Perth, WA, Australia.
authors.forename: Grant
authors.initials: G
authors.lastname: Morahan
authors.affiliation: Biotechnology and Drug Development Research Laboratory, School of Pharmacy, Curtin Health Innovation Research Institute (CHIRI), Curtin University, Perth, WA, Australia. hani.al-salami@curtin.edu.au.
authors.forename: Hani
authors.initials: H
authors.lastname: Al-Salami
mesh_terms: D000464:Alginates / Q000737:chemistry; D000818:Animals; D002214:Capsules; D002470:Cell Survival; D016207:Cytokines / Q000032:analysis / Q000378:metabolism*; D004339:Drug Compounding; D005260:Female; D005947:Glucose / Q000378:metabolism; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D007328:Insulin / Q000032:analysis / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D007515:Islets of Langerhans / Q000378:metabolism; D008822:Mice, Transgenic; D008853:Microscopy; D010455:Peptides / Q000737:chemistry; D000072760:Single Molecule Imaging / Q000379:methods*; D014580:Ursodeoxycholic Acid / Q000737:chemistry
chemical_list: D000464:Alginates; D002214:Capsules; D016207:Cytokines; D006603:Hexuronic Acids; D007328:Insulin; D053583:Interleukin-1beta; D010455:Peptides; C008973:polyornithine; D014580:Ursodeoxycholic Acid; D020723:Glucuronic Acid; D005947:Glucose
keywords: Cytokines; Microencapsulation; Microscopy; RIP-DsRED pancreatic β-cells; Ursodeoxycholic acid