Cryopreservation of Primarily Isolated Porcine Hepatocytes with UW Solution

Autor: Takemi Kunieda, Masanobu Maruyama, Teru Okitsu, Norikuni Shibata, Michihiko Takesue, Toshinori Totsugawa, Yoshikazu Kosaka, Takashi Arata, Kazuya Kobayashi, Hideaki Ikeda, Mizuko Oshita, Shuhei Nakaji, Kenji Ohmoto, Shinichiro Yamamoto, Yuzuru Kurabayashi, Makoto Kodama, Noriaki Tanaka, Naoya Kobayashi M.D., PH.D.
Jazyk: angličtina
Rok vydání: 2003
Předmět:
Zdroj: Cell Transplantation, Vol 12 (2003)
Druh dokumentu: article
ISSN: 0963-6897
1555-3892
00000000
DOI: 10.3727/000000003108747217
Popis: Development of liver-targeted cell therapies, such as hepatocyte transplantation and bioartificial livers, requires a large amount of functional hepatocytes as needed. To achieve this development, establishing an excellent cryopreservation method of hepatocytes is an extremely important issue. Therefore, we performed a comparative review of cryoprotective effects of various cryopreservation solutions using primarily isolated porcine hepatocytes. Porcine hepatocytes were isolated with a four-step dispase and collagenase perfusion method. The obtained hepatocytes with the initial viabilities of 76%, 84%, and 96% were assigned to the following four groups for cryopreservation at −80°C: Dulbecco's modified Eagle's medium (DMEM) + 10% fetal bovine serum (FBS) + 12% dimethyl sulfoxide (DMSO) (group A), University of Wisconsin (UW) solution + 12% DMSO (group B), Cell Banker 1 (group C), and Cell Banker 2 (group D). The hepatocytes in each group were thawed at 3 days, 10 days, and 5 months of cryopreservation and subjected to comparative analyses, including viability, plating efficiency, LDH release, ammonia removal test, and lentiviral gene transfer. These parameters were the most favorable in the hepatocytes cryopreserved with UW solution. Approximately 5% of thawed cryopreserved porcine hepatocytes expressed LacZ activity after lentiviral transduction. Intrasplenic transplantation of UW solution-cryopreserved hepatocytes improved the survival of rats treated with D-galactosamine. UW solution maintained the functions of cryopreserved porcine hepatocytes.
Databáze: Directory of Open Access Journals
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