Cryopreservation of Primarily Isolated Porcine Hepatocytes with UW Solution
| Autor: | Teru Okitsu, Hideaki Ikeda, Noriaki Tanaka, Shinichiro Yamamoto, N Kobayashi, Masanobu Maruyama, Toshinori Totsugawa, Takemi Kunieda, Kazuya Kobayashi, Makoto Kodama, Takashi Arata, Norikuni Shibata, Mizuko Oshita, Shuhei Nakaji, Kenji Ohmoto, Yoshikazu Kosaka, Yuzuru Kurabayashi, Michihiko Takesue |
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| Rok vydání: | 2003 |
| Předmět: |
Male
0301 basic medicine Adenosine Plating efficiency Cell Survival Cell Transplantation Allopurinol Genetic Vectors Organ Preservation Solutions Sus scrofa Transplantation Heterologous Cell Culture Techniques Biomedical Engineering lcsh:Medicine Cell Separation Cryopreservation Andrology 03 medical and health sciences chemistry.chemical_compound Cryoprotective Agents Raffinose 0302 clinical medicine Ammonia Transduction Genetic Dispase medicine Animals Insulin Viaspan Rats Wistar Transplantation L-Lactate Dehydrogenase Chemistry Dimethyl sulfoxide Liver Diseases lcsh:R Cell Biology Glutathione Liver Transplantation Rats 030104 developmental biology medicine.anatomical_structure Lac Operon Hepatocyte Hepatocytes 030217 neurology & neurosurgery Fetal bovine serum |
| Zdroj: | Cell Transplantation, Vol 12 (2003) |
| ISSN: | 1555-3892 0963-6897 |
| 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: | OpenAIRE |
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