title: Assessment of six different collagenase-based methods to isolate feline pancreatic islets.
abstract: Isolation of pancreatic islets is necessary to study the molecular mechanisms underlying beta-cell demise in diabetic cats. Six collagenase-based methods of isolation were compared in 10 cat pancreata, including single and double course of collagenase, followed or not by Ficoll centrifugation or accutase, and collagenase plus accutase. Morphometric analysis was performed to measure the relative area of islet and exocrine tissue. Islet specific mRNA transcripts were quantified in isolates by real-time PCR. The single and double course of collagenase digestion was successful in each cat and provided similar islet-to-exocrine tissue ratio. Quantities of insulin mRNA did not differ between the two methods. However, on histological examination either method yielded only approximately 2% of pure islets. The other methods provided disrupted islets or insufficient samples in 1-7 cats. Although pancreas digestion with single and double course of collagenase was superior, further studies are needed to improve islet isolation in cats.
authors.affiliation: Clinic for Small Animal Internal Medicine, Vetsuisse Faculty, University of Zürich, Winterthurerstr. 260, 8057 Zürich, Switzerland. ezini@vetclinics.uzh.ch
authors.forename: Eric
authors.initials: E
authors.lastname: Zini
authors.forename: Marco
authors.initials: M
authors.lastname: Franchini
authors.forename: Franco
authors.initials: F
authors.lastname: Guscetti
authors.forename: Melania
authors.initials: M
authors.lastname: Osto
authors.forename: Karin
authors.initials: K
authors.lastname: Kaufmann
authors.forename: Mathias
authors.initials: M
authors.lastname: Ackermann
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Lutz
authors.forename: Claudia E
authors.initials: CE
authors.lastname: Reusch
mesh_terms: D000818:Animals; D002415:Cats; D018929:Cell Culture Techniques / Q000662:veterinary; D017364:Collagenases / Q000378:metabolism*; D007515:Islets of Langerhans / Q000166:cytology*; D010179:Pancreas / Q000201:enzymology / Q000378:metabolism
chemical_list: D017364:Collagenases