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However there are limitations to the supply of human liver from surgical resection. Therefore, the efficient use of a limited liver supply is required. For the purpose it is extremely important to establish efficient techniques for the cell isolation and cold-preservation of isolated liver cells. As for the isolation of hepatocytes from surgically removed pig livers, dispase perfusion followed by collagenase digestion should yield hepatocytes with more than 90% viability. Cold-preservation for eight hours using isolated pig hepatocytes in the following preservation media was compared ; 1) University of Wiconsin (UW) solution with ascorbic acid 2 glucocide (AA2G) 2) University of Wisconsin (UW) solution 3) 100% fetal bovine serum 4) Dulbecco\u0027s Modified Eagle\u0027s Medium supplemented with 10% fetal bovine serum With the UW solution, viability, plating efficiency and ammonium clearance were well preserved after eight hours of cold-preervation. 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ブタ分離肝細胞の冷保存に関する基礎的検討
https://kwmed.repo.nii.ac.jp/records/1267
https://kwmed.repo.nii.ac.jp/records/126720bef1b3-9fea-42f3-af98-47fcc5612915
名前 / ファイル | ライセンス | アクション |
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KJ00001725526.pdf (982.4 kB)
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Item type | [ELS]学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2017-01-23 | |||||
タイトル | ||||||
言語 | ja | |||||
タイトル | ブタ分離肝細胞の冷保存に関する基礎的検討 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Cold-Preservation of Primariy Isolated Pig Hepatocytes : Fundamental Study Concerning Optimal Medium | |||||
言語 | ||||||
言語 | jpn | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
武居, 道彦
× 武居, 道彦× TAKESUE, Michihiko |
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著者所属(日) | ||||||
ja | ||||||
川崎医科大学内科消化器I部門 | ||||||
著者所属(英) | ||||||
en | ||||||
Division of Gastroenteroloy, Department of Medicine, Kawasaki Medical School | ||||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Cold preservation | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Isolated porcine liver cells | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Ascorbic acid-2 glucocide | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | University of Wisconsin solution | |||||
記事種別(日) | ||||||
内容記述タイプ | Other | |||||
内容記述 | 原著 | |||||
言語 | ja | |||||
抄録(日) | ||||||
ja | ||||||
肝再生医療の資源として正常ヒト肝細胞の確保が理想であるが,深刻なドナー不足の現状である.この問題を解決するためには,限られた提供臓器をいかに有効利用するかがきわめて重要である.それには,肝細胞の効率的な分離と効果的な細胞の冷保存法の確立が必要である.外科的に切除したブタ肝からの肝細胞の分離は,collagenaseとdispaseを組み合わせることにより90%以上のviabilityを有する肝細胞を得る事が可能であった.さらに当該細胞を用いて8時間冷保存に耐えうるか否かについて各種保存液の比較検討を行った.冷保存液として,(1) University of Wisconsin solution(以下UW)液,(2) UW液にvitamin Cの誘導体である2-O-a-D-Glucopyranosyl-L-Ascorbic Acid(以下ascorbic acid-2 glucoside ; AA2G) 100μg./mlを添加した保存液,(3)100% fetal bovine serum(以下,FBS), (4) Dulbecco's Modified Eagle's Medium(以下,DMEM)に10% FBSを入れた保存液の4種類を検討した.UW液を使用することで,有意に8時間冷保存ブタ肝細胞のviability, plating efficiency,アンモニア代謝能が維持された.さらに,UW液に強力な抗酸化剤であるvitamin C配糖体であるAA2Gを組み合わせることで更に効果的に冷保存肝細胞の機能を維持することができた.その理由としてAA2G添加UW液では活性型caspase-3の誘導が抑制され,細胞内ATP量が有意に保持されたためであると考えられる.当該結果から,UW液+AA2Gは冷保存に最も適した保存液であり,臨床応用への有効性が示唆された. | ||||||
抄録(英) | ||||||
en | ||||||
As a source for liver-targeted regeneration medicine, a supply of normal human hepatocytes is essential. However there are limitations to the supply of human liver from surgical resection. Therefore, the efficient use of a limited liver supply is required. For the purpose it is extremely important to establish efficient techniques for the cell isolation and cold-preservation of isolated liver cells. As for the isolation of hepatocytes from surgically removed pig livers, dispase perfusion followed by collagenase digestion should yield hepatocytes with more than 90% viability. Cold-preservation for eight hours using isolated pig hepatocytes in the following preservation media was compared ; 1) University of Wiconsin (UW) solution with ascorbic acid 2 glucocide (AA2G) 2) University of Wisconsin (UW) solution 3) 100% fetal bovine serum 4) Dulbecco's Modified Eagle's Medium supplemented with 10% fetal bovine serum With the UW solution, viability, plating efficiency and ammonium clearance were well preserved after eight hours of cold-preervation. UW + AA2G solution yielded the best preservation effect. The use of AA2G inhibited activation of caspase-3, resulting in well-preserved cellular ATP levels. Based on the above results, the UW + AA2G solution appeared to be most suitable for cold-preservation of pig hepatocytes. Clininal application for the cold-preservation of human hepatocytes can be expected. | ||||||
書誌情報 |
ja : 川崎医学会誌 en : Kawasaki medical journal 巻 28, 号 3, p. 199-208, 発行日 2002 |
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URL | ||||||
識別子 | http://igakkai.kms-igakkai.com/wp/wp-content/uploads/2002/KMJ28(3)199-208.2002.pdf | |||||
識別子タイプ | URI | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | https://doi.org/10.11482/KMJ28(3)199-208.2002.pdf | |||||
ISSN | ||||||
収録物識別子タイプ | PISSN | |||||
収録物識別子 | 0386-5924 | |||||
ISSN | ||||||
収録物識別子タイプ | EISSN | |||||
収録物識別子 | 2758-089X | |||||
雑誌書誌ID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AN00045593 | |||||
雑誌書誌ID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AN12940574 | |||||
著者版フラグ | ||||||
出版タイプ | VoR |