Maintenance of cold-preserved porcine hepatocyte function with UW solution and ascorbic acid-2 glucoside

Michihiko Takesue, Masanobu Maruyama, Norikuni Shibata, Takemi Kunieda, Teru Okitsu, Masakiyo Sakaguchi, Toshinori Totsugawa, Yoshikazu Kosaka, Akira Arata, Hideaki Ikeda, Junji Matsuoka, Toshie Oyama, Makoto Kodama, Kenji Ohmoto, Shinichiro Yamamoto, Yuzuru Kurabayashi, Itaru Yamamoto, Noriaki Tanaka, Naoya Kobayashi

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)599-606
Number of pages8
JournalCell Transplantation
Volume12
Issue number6
DOIs
Publication statusPublished - Jan 1 2003

Keywords

  • Ascorbic acid-2 glucoside
  • Cold preservation
  • Hepatocytes
  • University of Wisconsin solution

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

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    Takesue, M., Maruyama, M., Shibata, N., Kunieda, T., Okitsu, T., Sakaguchi, M., Totsugawa, T., Kosaka, Y., Arata, A., Ikeda, H., Matsuoka, J., Oyama, T., Kodama, M., Ohmoto, K., Yamamoto, S., Kurabayashi, Y., Yamamoto, I., Tanaka, N., & Kobayashi, N. (2003). Maintenance of cold-preserved porcine hepatocyte function with UW solution and ascorbic acid-2 glucoside. Cell Transplantation, 12(6), 599-606. https://doi.org/10.3727/000000003108747208