title: Effective islet isolation method with extremely high islet yields from adult pigs.
abstract: Achieving good islet isolation is one of the most important factors for successful islet transplantation. Porcine pancreas is suitable for islet isolation research due to its anatomical and physiological similarities to human pancreas. In this study, we evaluated a new porcine islet isolation method designed to maximize islet yield and compared it with our previous open pan method and the standard method using a Ricordi chamber (Ricordi method). We performed 15 porcine islet isolations, five each with the new method, the open pan method, and the Ricordi method. The new method features several important improvements. Pancreata remain uncut and are kept intact during collagenase intraductal injection, a large filtration chamber to handle whole pancreata, low concentration of collagenase (Liberase HI) for digestion, and large plastic containers for large-scale islet purification. All isolated islets were assessed for yield, purity, viability and in vitro function. Islets isolated with this new method were transplanted under the kidney capsules of SCID mice with chemically induced diabetes for in vivo functional assessment (n = 8). With the new method, we obtained on average more than 1,000,000 islet equivalents (IE) (1,236,266 +/- 213,486 IE) (mean +/- SE) before purification and 800,000 IE (879,815 +/- 222,729 IE) after purification from one adult pig. Islet yield per pancreas was significantly higher compared with our previous open pan method (30,666 +/- 11,532 IE, p < 0.01) and the Ricordi method (317,073 +/- 86,093 IE, p < 0.05). All mice, transplanted with 1000 islets from the new method, returned to normoglycemia within 4 days after transplantation. Our new method makes it possible to obtain extremely high porcine islet yield with good function. It should produce useful information for human islet isolation and transplantation, and might be applied to single donor clinical xenogeneic transplantation.
authors.affiliation: Department of Transplantation and Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Japan.
authors.forename: Yukihide
authors.initials: Y
authors.lastname: Yonekawa
authors.forename: Shinichi
authors.initials: S
authors.lastname: Matsumoto
authors.forename: Teru
authors.initials: T
authors.lastname: Okitsu
authors.forename: Takashi
authors.initials: T
authors.lastname: Arata
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Iwanaga
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Noguchi
authors.forename: Hideo
authors.initials: H
authors.lastname: Nagata
authors.forename: John J
authors.initials: JJ
authors.lastname: O'Neil
authors.forename: Koichi
authors.initials: K
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D049628:Body Size; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D017364:Collagenases; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000601:surgery; D005951:Glucose Tolerance Test; D007515:Islets of Langerhans / Q000166:cytology* / Q000473:pathology; D016381:Islets of Langerhans Transplantation; D051379:Mice; D016513:Mice, SCID; D013552:Swine; D014183:Transplantation, Heterologous
chemical_list: D001786:Blood Glucose; D017364:Collagenases