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ID 52864
フルテキストURL
著者
Arata, T Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Okitsu, T Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Fukazawa, T Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Ikeda, H Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Kobayashi, K Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Yong, C Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Kosaka, Y Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Narushima, M Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Matsuoka, J Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
Yamamoto, I Department of Immunochemistry, Faculty of Pharmaceutical Sciences, Okayama University
Tanaka, N Department of Immunochemistry, Faculty of Pharmaceutical Sciences, Okayama University
Lakey, JRT Univ Alberta, Surg Med Res Inst, Clin Islet Transplantat Program
Kobayashi, N Department of Surgery, Okayama University Graduate School of Medicine and Dentistry
抄録
Normal human islet cells are an ideal source for pancreas-targeted cell therapies, but the availability of human donor pancreata for islet isolation is severely limited. To effectively utilize such scarce donor organs for cell therapies, it is crucial to develop an excellent isolation, effective cryopreservation, and efficient gene transfer techniques for the transportation of isolated cells. In the present study, we investigate the effect of University of Wisconsin (UW) solution and ascorbic acid-2 glucoside (AA2G) on the cryopreservation of human islets. We also evaluate the gene transfer efficiency of a lentiviral vector expressing the E. coli LacZ gene, Lt-NLS/LacZ, in human islets. Human islets were isolated with a standard digestion method at the University of Alberta. Isolated islets were transported to Japan for 40 h and then subjected to cryopreservation experiments. The following preservation solutions were tested: UW solution with 100 mug/mL of AA2G, UW solution, 100% fetal bovine serum (FBS), and CMRL supplemented with 10% FBS. Following three months of cryopreservation, the islets were thawed and analyzed for viability, glucose-sensitive insulin secretion, proinsulin gene expression profile, and in vivo engraftment. The islets were also subjected to monolayer formation with 804G-cell-line-derived extracellular matrix (ECM), followed by Lt-NLS/LacZ transduction. The viability, morphology, glucose-sensitive insulin secretion, proinsulin gene expression, and monolayer formation efficiency of the thawed cryopreserved islets are significantly better maintained by the use of UW solution. When AA2G (100 mug/mL) is combined with UW, such parameters are further improved. The adequate engraftment of UW + AA2G-cryopreserved human islets is achieved in the liver of nude mice. Efficient Lt-NLS/LacZ transduction is identified in monolayered islets cryopreserved with UW solution with AA2G. The present work demonstrates that the combination of UW solution with AA2G (100 mug/mL) would be a useful cryopreservation means for human islets. Human islets monolayer-cultured with 804G-derived ECM are efficiently transduced with a lentiviral vector Lt-NLS/LacZ.
キーワード
islets
University of Wisconsin solution
ascorbic acid-2 glucoside
cryopreservation
stimulation index
発行日
2004-06
出版物タイトル
Artificial Organs
28巻
6号
開始ページ
529
終了ページ
536
資料タイプ
学術雑誌論文
関連URL
http://ousar.lib.okayama-u.ac.jp/metadata/52835
言語
English
OAI-PMH Set
岡山大学
論文のバージョン
author
査読
有り
DOI
Web of Sience KeyUT