title: An effective purification method using large bottles for human pancreatic islet isolation.
abstract: The purification process is one of the most difficult procedures in pancreatic islet isolation. It was demonstrated that the standard purification method using a COBE 2991 cell processor with Ficoll density gradient solution harmed islets mechanically by high shear force. We reported that purification using large bottles with a lower viscosity gradient solution could improve the efficacy of porcine islet purification. In this study, we examined whether the new bottle purification method could improve the purification of human islets. Nine human pancreata from brain-dead donors were used. After pancreas digestion, the digested tissue was divided into three groups. Each group was purified by continuous density gradient using ET-Kyoto and iodixanol gradient solution with either the standard COBE method (COBE group) or the top loading (top group) or bottom loading (bottom group) bottle purification methods. Islet yield, purity, recovery rate after purification, and in vitro and in vivo viability were compared. Islet yield per pancreas weight (IE/g) and the recovery rate in the top group were significantly higher than in the COBE and bottom groups. Furthermore, the average size of purified islets in the top group was significantly larger than in the COBE group, which indicated that the bottle method could reduce the shear force to the islets. In vivo viability was also significantly higher in the top group compared with the COBE group. In conclusion, the top-loading bottle method could improve the quality and quantity of human islets after purification.
authors.affiliation: Baylor University Medical Center, Dallas, TX, USA.unimogura@hotmail.co.jp
authors.forename: Masayuki
authors.initials: M
authors.lastname: Shimoda
authors.forename: Takeshi
authors.initials: T
authors.lastname: Itoh
authors.forename: Shuichi
authors.initials: S
authors.lastname: Iwahashi
authors.forename: Morihito
authors.initials: M
authors.lastname: Takita
authors.forename: Koji
authors.initials: K
authors.lastname: Sugimoto
authors.forename: Mazhar A
authors.initials: MA
authors.lastname: Kanak
authors.forename: Daisuke
authors.initials: D
authors.lastname: Chujo
authors.forename: Bashoo
authors.initials: B
authors.lastname: Naziruddin
authors.forename: Marlon F
authors.initials: MF
authors.lastname: Levy
authors.forename: Paul A
authors.initials: PA
authors.lastname: Grayburn
authors.forename: Shinichi
authors.initials: S
authors.lastname: Matsumoto
mesh_terms: D000328:Adult; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002499:Centrifugation, Density Gradient / Q000295:instrumentation / Q000379:methods*; D003921:Diabetes Mellitus, Experimental / Q000601:surgery; D005260:Female; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology*; D016381:Islets of Langerhans Transplantation; D053208:Kaplan-Meier Estimate; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D010179:Pancreas / Q000166:cytology*
chemical_list: D001786:Blood Glucose