title: Viable purified islets of Langerhans from collagenase-perfused human pancreas.
abstract: Human islets of Langerhans were isolated with the principles of collagenase perfusion via the pancreatic duct and gentle dissociation of tissue. The number of islets released was 161 x 10(3), distributed as 76 x 10(3) large (greater than 100-micron) and 85 x 10(3) small (less than 100-micron) islets. Recovery after Ficoll-gradient purification was 61% for the large islets and 42% for the small islets. The final islet volume was 240 microliter, with purity of 70-90% (large islets) and 20-40% (small islets). Perifusion with glucose elicited a biphasic release of insulin, with the response rising sixfold from basal secretion. Implantation of pure islets under the kidney capsule of normal or streptozocin-induced diabetic nude mice resulted in human C-peptide secretion and partial or complete reversal of hyperglycemia, confirmed by histological recovery. The data show that these methods provide large quantities of viable purified human islets.
authors.affiliation: Department of Surgery, University of Alberta, Edmonton, Canada.
authors.forename: G L
authors.initials: GL
authors.lastname: Warnock
authors.forename: D K
authors.initials: DK
authors.lastname: Ellis
authors.forename: M
authors.initials: M
authors.lastname: Cattral
authors.forename: D
authors.initials: D
authors.lastname: Untch
authors.forename: N M
authors.initials: NM
authors.lastname: Kneteman
authors.forename: R V
authors.initials: RV
authors.lastname: Rajotte
mesh_terms: D000328:Adult; D000818:Animals; D002096:C-Peptide / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D003012:Microbial Collagenase / Q000378:metabolism*; D010179:Pancreas / Q000166:cytology*
chemical_list: D002096:C-Peptide; D003012:Microbial Collagenase