Maintenance of Cold-Preserved Porcine Hepatocyte Function with UW Solution and Ascorbic Acid-2 Glucoside

Cell Transplantation

Article

Yuzuru Kurabayashi¹  Masakiyo Sakaguchi²  Itaru Yamamoto³  Noriaki Tanaka⁴  Akira Arata⁴  Yoshikazu Kosaka⁴  Teru Okitsu⁴  Toshinori Totsugawa⁴  Hideaki Ikeda⁴  Naoya Kobayashi⁴  Junji Matsuoka⁴  Masanobu Maruyama⁴  Takemi Kunieda⁵  Norikuni Shibata⁵  Shinichiro Yamamoto⁵  Kenji Ohmoto⁵  Michihiko Takesue⁵  Makoto Kodama⁶  Toshie Oyama⁶ 

[1] Animal Center for Medical Research, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan;Department of Cell Biology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan;Department of Immunochemistry, Faculty of Pharmaceutical Sciences, Okayama University, 1-1-1 Tsushima-naka, Okayama 700-8530, Japan;Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan;Division of Gastroenterology, Department of Internal Medicine, Kawasaki Medical School, 577 Matsushima, Kurashiki 701-0192 Japan;Tissue Engineering Research Center, The National Institute of Advanced Industrial Science and Technology, 1-1-4 Higashi, Tsukubashi, Ibaragi 305-8562, Japan;

关键词: Hepatocytes;    University of Wisconsin solution;    Ascorbic acid-2 glucoside;    Cold preservation;   

DOI  :  10.3727/000000003108747208

来源: Sage Journals

【 摘 要 】

Normal human hepatocytes are an ideal source of liver-targeted cell therapies, such as hepatocyte transplantation and bioartificial livers, but availability of human donor livers for liver cell isolation is severely limited. To effectively utilize scarce donor organs for cell therapies, it is of extreme importance to establish an efficient isolation technique and an effective cold preservation solution for transportation of isolated cells. A lateral segment of the liver was surgically resected from pigs weighing 10 kg and a four-step collagenase and dispase digestion was conducted. Isolated hepatocytes were subjected to 8-h cold storage on ice. The following preservation solutions were tested: 1) University of Wisconsin (UW) solution, 2) UW with 100 μg/ml of ascorbic acid-2 glucoside (AA2G), 3) 100% fetal bovine serum (FBS), and 4) Dulbecco's modified Eagle's medium (DMEM) supplemented with 100% FBS. The mean viability of porcine hepatocytes was 95.5 ± 2.5% when isolated in three independent experiments. Viability, plating efficiency, membrane stability, and ammonia metabolic capacity of cold-preserved hepatocytes were significantly better maintained by the use of UW solution. When AA2G (100 μg/ml) was combined with UW solution, such parameters were further improved. It was explained by inhibition of caspase-3 activation and retention of ATP at high levels of hepatocytes preserved with UW solution containing AA2G. The present work demonstrates that a combination of UW solution with AA2G (100 μg/ml) would be a useful cold preservation means for the development of cell therapies.

【 授权许可】

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