title: Storage of human pancreatic digest in University of Wisconsin solution significantly improves subsequent islet purification.
abstract: Density-gradient purification of human pancreatic islets from the collagenase-digested pancreas relies on the exocrine tissue being denser than the islets. Cold storage of the pancreas before and after digestion causes cell swelling, which can decrease the density of pancreatic exocrine tissue and adversely affect subsequent purification. Using 14 human pancreata (seven perfused in situ with hyperosmolar citrate (HOC) and seven with University of Wisconsin solution (UW)), it is shown that storage of the pancreatic digest in UW significantly increases the density of pancreatic exocrine tissue compared with storage in minimal essential medium (MEM) (P = 0.009). This results in an improvement in islet purity (P = 0.036) for HOC- but not UW-perfused pancreata. Storage in UW for 1 h not only prevented the deterioration that occurred in MEM, but resulted in an improvement in islet purity for five of the seven HOC-perfused pancreata. Most pancreata in the UK are perfused with HOC, but storage of the digest in UW results in significantly better islet purity and, when islets cannot be purified immediately, a period of storage will often improve separation and allow islets to be purified.
authors.affiliation: Department of Surgery, Leicester Royal Infirmary, UK.
authors.forename: G S
authors.initials: GS
authors.lastname: Robertson
authors.forename: D
authors.initials: D
authors.lastname: Chadwick
authors.forename: H
authors.initials: H
authors.lastname: Contractor
authors.forename: S
authors.initials: S
authors.lastname: Rose
authors.forename: R
authors.initials: R
authors.lastname: Chamberlain
authors.forename: H
authors.initials: H
authors.lastname: Clayton
authors.forename: P R
authors.initials: PR
authors.lastname: Bell
authors.forename: R F
authors.initials: RF
authors.lastname: James
authors.forename: N J
authors.initials: NJ
authors.lastname: London
mesh_terms: D000241:Adenosine; D000493:Allopurinol; D005978:Glutathione; D006801:Humans; D007328:Insulin; D007515:Islets of Langerhans; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D009994:Osmolar Concentration; D011887:Raffinose; D012996:Solutions
chemical_list: D007328:Insulin; D019188:Organ Preservation Solutions; D012996:Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose