title: Comparative study on biologic and immunologic characteristics of the pancreas islet cell between 24 degrees C and 37 degrees C culture in the rat.
abstract: The aim of this study was to investigate the effect of culture at 24 degrees C on cell viability, cellular function, immunogenicity, and cytokine profiles of rat pancreatic islets. Pancreatic islets were isolated from Lewis rats and cultured at either 24 degrees C or 37 degrees C for 14 days. Islet recovery was counted as islet equivalents; islet viability was examined with fluorescent vital staining. Islet function was measured with a glucose stimulation test. Annexin V, and MHC class I and II expression were measured using flow cytometric assay for apoptosis and immunogenicity, respectively. Lymphocyte cell proliferation was examined with WST-1 proliferation assay. Cytokine profiles were analyzed with quantitative real time RT-PCR. All these parameters were measured on 1, 3, 5, 7 and 14 culture days after islet isolation. Islet recovery was higher in islets cultured at 24 degrees C than 37 degrees C without a change in viability. Insulin secretion after glucose stimulation was more effective in 24 degrees C culture conditions. Decreased apoptotic cell death was demonstrated in 24 degrees C cultured islets. Both MHC class I and II expression on islets and lymphocyte proliferation upon coculture with islets were less prominent in 24 degrees C cultured islets. TNF-alpha expression was lower in islets cultured at 24 degrees C than in islets cultured at 37 degrees C. Both IL-1beta and IL-10 cytokine expressions were similar under both culture conditions. This study demonstrated that cell recovery and function are increased in islets cultured at 24 degrees C than those at 37 degrees C with decreased antigenicity and proinflammatory cytokine expression.
authors.affiliation: Department of Surgery, Ulsan University College of Medicine and Asan Medical Center, 388-1 Poongnab-dong, Songpa-gu, Seoul 138-736, Korea. drksc@amc.seoul.kr
authors.forename: S C
authors.initials: SC
authors.lastname: Kim
authors.forename: D J
authors.initials: DJ
authors.lastname: Han
authors.forename: I H
authors.initials: IH
authors.lastname: Kim
authors.forename: K O
authors.initials: KO
authors.lastname: Woo
authors.forename: Y M
authors.initials: YM
authors.lastname: We
authors.forename: S Y
authors.initials: SY
authors.lastname: Kang
authors.forename: J H
authors.initials: JH
authors.lastname: Back
authors.forename: Y H
authors.initials: YH
authors.lastname: Kim
authors.forename: J H
authors.initials: JH
authors.lastname: Kim
authors.forename: D G
authors.initials: DG
authors.lastname: Lim
mesh_terms: D000818:Animals; D017209:Apoptosis; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002469:Cell Separation; D016207:Cytokines / Q000235:genetics; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D011917:Rats, Inbred Lew; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature
chemical_list: D016207:Cytokines; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D007328:Insulin