title: Subcutaneous xenotransplantation of hybrid artificial pancreas encapsulating pancreatic B cell line (MIN6): functional and histological study.
abstract: The biohybrid artificial pancreas is designed to enclose pancreatic tissues with a selectively permeable membrane that immunoisolates the graft from the host immune system, allowing those endocrine tissues to survive and control glucose metabolism for an extended period of time. The pancreatic B cell line MIN6 is established from a pancreas B cell tumor occurring in transgenic mice harbouring the human insulin promoter gene connected to the SV40 T-antigen hybrid gene. It has been proven that glucose-stimulated insulin secretion in MIN6 cells retains a concentration-dependent response similar to that of normal islets. In this study, we performed the histological and functional examination of three-layer microbeads employing MIN6 cells after subcutaneous xenotransplantation to evaluate this device as bioartificial pancreas. MIN6 cells were microencapsulated in three-layer microbeads formulated with agarose, polystyrene sulfonic acid, polybrene, and carboxymethyl cellulose. Microbeads were xenogenically implanted in the subcutaneous tissue of the back of Lewis rats with streptozotocin-induced diabetes. One week after implantation, microbeads were retrieved and cultured for 24 h before the static incubation. There was no evidence of adhesion to the graft and the fibrosis in the transplantation site as determined by gross visual inspection. Microscopic examination demonstrated that retrieved microbeads maintained normal shape, containing intact MIN6 cells. Histological study showed that these MIN6 cells in the microbeads appeared to be viable without cellular infiltration within or around the microbeads. Immunohistochemical analysis of the microbeads clearly revealed the intense staining of insulin in the cytoplasm of encapsulated MIN6 cells. Insulin productivity of MIN6 cells in the microbeads is strongly suggested to be preserved. In response to 16.7 mM glucose stimulation, static incubation of microbeads 1 wk after implantation caused the 2.3 times increase in insulin secretion seen after 3.3 mM glucose stimulation (84.3 +/- 10.0 vs. 37.4 +/- 10.7 microU/3 x 10(6) cells/hr, n = 5 each, p < 0.01). This study demonstrates that three-layer microbeads encapsulating MIN6 cells retain excellent biocompatibility and maintain good insulin secretion even after subcutaneous xenotransplantation, suggesting the possible future clinical application of this unique bioartificial pancreas to subcutaneous xenotransplantation.
authors.affiliation: First Department of Surgery, Faculty of Medicine, Kyoto University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kawakami
authors.forename: K
authors.initials: K
authors.lastname: Inoue
authors.forename: H
authors.initials: H
authors.lastname: Hayashi
authors.forename: W J
authors.initials: WJ
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Setoyama
authors.forename: Y J
authors.initials: YJ
authors.lastname: Gu
authors.forename: M
authors.initials: M
authors.lastname: Imamura
authors.forename: H
authors.initials: H
authors.lastname: Iwata
authors.forename: Y
authors.initials: Y
authors.lastname: Ikada
authors.forename: M
authors.initials: M
authors.lastname: Nozawa
authors.forename: J
authors.initials: J
authors.lastname: Miyazaki
mesh_terms: D000818:Animals; D002214:Capsules; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D016381:Islets of Langerhans Transplantation; D008297:Male; D051379:Mice; D009368:Neoplasm Transplantation; D019397:Pancreas, Artificial; D051381:Rats; D011917:Rats, Inbred Lew; D012867:Skin; D014183:Transplantation, Heterologous; D016042:Transplantation, Heterotopic; D014407:Tumor Cells, Cultured
chemical_list: D002214:Capsules; D007328:Insulin; D005947:Glucose