title: A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC).
abstract: BACKGROUND
  N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs.
  
  
  METHODS
  NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. The extract was proteolyzed by treatment with a chymotrypsin and trypsin mixture and further digested with glycoamidase A to release the N-glycans. After the removal of the peptide materials, the reducing ends of the N-glycans were derivatized with 2-aminopyridine. This mixture was applied to DEAE, amide and ODS columns. PA-oligosaccharides were also subjected to MALDI TOF-MS analysis.
  
  
  RESULTS
  The NPCC glycans were comprised of 14 neutral, 5 mono-sialyl and 5 di-sialyl glycans. As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. Moreover, most of the structures could be clearly identified in this study.
  
  
  CONCLUSIONS
  The data herein will be helpful for future studies of the glycoantigen associated with NPCC.
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: miyagawa@orgtrp.med.osaka-u.ac.jp.
authors.forename: Shuji
authors.initials: S
authors.lastname: Miyagawa
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Maeda
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Eguchi
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Takuji
authors.initials: T
authors.lastname: Kawamura
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kashiyama
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Takehisa
authors.initials: T
authors.lastname: Ueno
authors.affiliation: Division of Organ Transplantation (E9), Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Usui
authors.affiliation: Medical & Biological Laboratories Co., Ltd., Sakae 4-5-3, Naka-ku, Nagoya, Aichi 460-0008, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kondo
authors.affiliation: Department of Life Science, Meiji University, Higashisanda 1-1-1, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nakano
authors.affiliation: Department of Life Science, Meiji University, Higashisanda 1-1-1, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Matsunari
authors.affiliation: Department of Life Science, Meiji University, Higashisanda 1-1-1, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
authors.forename: Masahito
authors.initials: M
authors.lastname: Watanabe
authors.affiliation: Department of Life Science, Meiji University, Higashisanda 1-1-1, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nagashima
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002851:Chromatography, High Pressure Liquid; D007515:Islets of Langerhans / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine
chemical_list: D011134:Polysaccharides
keywords: N-glycan; NPCC; Non-Gal antigen; Xenotransplantation