Development of an efficient preculture system of islets is ideal. Toward that goal, we constructed a human pancreatic islet-derived fibroblast cell line MNNK-1 for a source as a coculture system for freshly isolated islets to maintain islet functions. Human pancreatic islet cells were nucleofected with a plasmid vector pYK-1 expressing simian virus 40 large T antigen gene (SV40T) and hygromycin resistance gene (HygroR). One of the transduced cell lines, MNNK-1, was established and served as a feeder cell in the coculture for freshly isolated mouse, rat, and pig islets. Morphology, viability, and glucose-responding insulin secretion were analyzed in the coculture system. MNNK-1 cells were morphologically spindle shaped and were negative for pancreatic endocrine markers. MNNK-1 cells were positive for α-smooth muscle actin and collagen type I and produced fibroblast growth factor. Coculture of the mouse, rat, and pig islets with MNNK-1 cells maintained their viability and insulin secretion with glucose responsiveness. A human pancreatic islet-derived fibroblast cell line MNNK-1 was established. MNNK-1 cells were a useful means for maintaining morphology and insulin secretion of islets in the coculture system.
ArataT.; OkitsuT.; FukazawaT.; IkedaH.; KobayashiK.; YongC.; KosakaY.; NarushimaM.; MatsuokaJ.; YamamotoI.; TanakaN.; LakeyJ. R.; KobayashiN.Maintenance of glucose-sensitive insulin secretion of cryopreserved human islets with University of Wisconsin solution and ascorbic acid-2 glucoside.Artif. Organs28: 529–536; 2004.
3.
BaidalD. A.; FroudT.; FerreiraJ. V.; KhanA.; AlejandroR.; RicordiC.The bag method for islet cell infusion.Cell Transplant.12: 809–813; 2003.
4.
BarshesN. R.; LeeT.; GoodpastureS.; BrunicardiF. C.; AlejandroR.; RicordiC.; SoltesG.; BarthM.; HamiltonD.; GossJ. A.Achievement of insulin independence via pancreatic islet transplantation using a remote isolation center: A first-year review.Transplant. Proc.36: 1127–1129; 2004.
5.
BhatiaS. N.; BalisU. J.; YarmushM. L.; TonerM.Effect of cell-cell interactions in preservation of cellular phenotype: Cocultivation of hepatocytes and nonparenchymal cells.FASEB J.13: 1883–1900; 1999.
6.
BodnarA. G.; OuelletteM.; FrolkisM.; HoltS. E.; ChiuC. P.; MorinG. B.; HarleyC. B.; ShayJ. W.; LichtsteinerS.; WrightW. E.Extension of life-span by introduction of telomerase into normal human cells.Science279: 349–352; 1998.
7.
CelliG.; LaRochelleW. J.; MackemS.; SharpR.; MerlinoG.Soluble dominant-negative receptor uncovers essential roles for fibroblast growth factors in multi-organ induction and patterning.EMBO J.17: 1642–1655; 1998.
8.
CounterC. M.; AvilionA. A.; LeFeuvreC. E.; StewartN. G.; GreiderC. W.; HarleyC. B.; BacchettiS.Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity.EMBO J.11: 1921–1929; 1992.
9.
CounterC. M.; BotelhoF. M.; WangP.; HarleyC. B.; BacchettiS.Stablilization of short telomeres and telomerase activity accompany immortalization of Epastein-Barr virus-transformed human B lymphocytes.J. Virol.68: 3410–3414; 1994.
10.
GartnerH. V.; SeidlC.; LuckenbachC.; SchummG.; SeifriedE.; RitterH.; BultmannB.Genetic analysis of a sarcoma accidentally transplanted from a patient to a surgeon.N. Engl. J. Med.335: 1494–1496; 1996.
11.
GeorgesP.; MuirheadR. P.; WilliamsL.; HolmanS.; TabiinM. T.; DeanS. K.; TuchB. E.Comparison of size, viability, and function of fetal pig islet-like cell clusters after digestion using collagenase or liberase.Cell Transplant.11: 539–545; 2002.
12.
GossJ. A.; GoodpastorS. E.; BrunicardiF. C.; BarthM. H.; SoltesG. D.; GarberA. J.; HamiltonD. J.; AlejandroR.; RicordiC.Development of a human pancreatic islet-transplant program through a collaborative relationship with a remote islet-isolation center.Transplantation77: 462–466; 2004.
13.
HalvorsenT. L.; LeibowitzG.; LevineF.Telomerase activity is sufficient to allow transformed cells to escape from crisis.Mol. Cell. Biol.19: 1864–1870; 1999.
MatsumotoS.; QualleyS. A.; GoelS.; HagmanD. K.; SweetI. R.; PoitoutV.; StrongD. M.; RobertsonR. P.; ReemsJ. A.Effect of the two-layer (University of Wisconsin solution-perfluorochemical plus O2) method of pancreas preservation on human islet isolation as assessed by Edmonton isolation protocol.Transplantation74: 1414–1419; 2002.
21.
MinamiK.; YanoH.; MikiT.; NagashimaK.; WangC. Z.; TanakaH.; MiyazakiJ.; SeinoS.Insulin secretion and differential gene expression in glucose-responsive and -unresponsive MIN6 sublines.Am. J. Physiol. Endocrinol. Metab.279: E773–E781; 2000.
22.
MirallesF.; CzernichowP.; OzakiK.; ItohN.; ScharfmannR.Signaling through fibroblast growth factor receptor 2b plays a key role in the development of the exocrine pancreas.Proc. Natl. Acad. Sci. USA96: 6267–6272; 1999.
23.
MooltenF. L.; WellsJ. M.Curability of tumors bearing herpes thymidine kinase genes transferred by retroviral vectors.J. Natl. Cancer Inst.82: 297–300; 1990.
NoguchiH.; KobayashiN.; WestermanK. A.; SakaguchiM.; OkitsuT.; TotsugawaT.; WatanabeT.; MatsumuraT.; FujiwaraT.; UedaT.; MiyazakiM.; TanakaN.; LeboulchP.Controlled expansion of human endothelial cell populations by Cre/loxP-based reversible immortalization.Hum. Gene Ther.13: 321–334; 2002.
26.
OhtaniS.; KagawaS.; TangoY.; UmeokaT.; TokunagaN.; TsunemitsuY.; RothJ. A.; TayaY.; TanakaN.; FujiwaraT.Quantitative analysis of p53-trageted gene expression and visualization of p53 transcriptional activity following intratumoral administration of adenoviral p53 in vivo.Mol. Cancer Ther.3: 93–100; 2004.
27.
OkitsuT.; BartlettS. T.; HadleyG. A.; DrachenbergC. B.; FarneyA. C.Recurrent autoimmunity accelerates destruction of minor and major histoincompatible islet grafts in nonobese diabetic (NOD) mice.Am. J. Transplant.1: 138–145; 2001.
28.
ParadisK.; LangfordG.; LongZ.; HeneineW.; SandstromP.; SwitzerW. M.; ChapmanL. E.; LockeyC.; OnionsD.; OttoE.Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue.Science285: 1236–1241; 1999.
29.
RabinovitchA.; RussellT.; MintzD. H.Factors from fibroblasts promote pancreatic islet B cell survival in tissue culture.Diabetes28: 1108–1113; 1979.
30.
RayF. A.; KraemerP. M.Iterative chromosome mutation and selection as a mechanism of complete transformation of human diploid fibroblasts by SV40T antigen.Carcinogenesis14: 1511–1516; 1993.
31.
RicordiC.; GrayD. W.; HeringB. J.; KaufmanD. B.; WarnockG. L.; KnetemanN. M.; LakeS. P.; LondonN. J.; SocciC.; AlejandroR.Islet isolation assessment in man and large animals.Acta Diabetol. Lat.27: 185–195; 1990.
32.
ShapiroA. M.; LakeyJ. R.; RyanE. A.; KorbuttG. S.; TothE.; WarnockG. L.; KnetemanN. M.; RajotteR. V.Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen.N. Engl. J. Med.343: 230–238; 2000.
33.
ShapiroA. M.; RicordiC.; HeringB.Edmonton's islet success has indeed been replicated elsewhere.Lancet362: 1242; 2003.
34.
ShayJ. W.; WrightW. E.Quantitation of the frequency of immortalization of normal human diploid fibroblasts by SV40T large T-antigen.Exp. Cell Res.184: 109–118; 1989.
WestermanK. A.; LeboulchP.Reversible immortalization of mammalian cells mediated by retroviral transfer and site-specific recombination.Proc. Natl. Acad. Sci. USA93: 8971–8976; 1996.
37.
ZhuJ.; WangH.; BishopJ. M.; BlackburnE. H.Telomerase extends the lifespan of virus-transformed human cells without net telomere lengthening.Proc. Natl. Acad. Sci. USA96: 3723–3728; 1999.
