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Hydroxyethyl starch-based preservation solutions enhance gene therapy vector delivery under hypothermic conditions.
- Source :
-
Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society [Liver Transpl] 2008 Dec; Vol. 14 (12), pp. 1708-17. - Publication Year :
- 2008
-
Abstract
- Isolated liver perfusion offers a unique prospect for safe, effective targeting of gene therapies that can be directed against allograft rejection or recurrent diseases such as reinfection by hepatitis C virus (HCV). We aimed to examine the effect of organ preservation solutions on vector-based gene therapy delivery under hypothermic conditions. University of Wisconsin (UW) solution, histidine tryptophan ketoglutarate (HTK), EloHaes, sodium-poly(ethylene glycol)-UW solution [Institut Georges Lopez 1 solution (IGL-1)], and Dulbecco's modified Eagle's medium (DMEM) culture medium (control) were tested at 2 degrees C or 37 degrees C for lentiviral vector transduction efficiencies to the hepatoma cell line Huh-7 and primary human or mouse hepatocytes. Lentiviral vectors expressing short hairpin RNA were used to target HCV replication. With a potent short hairpin RNA vector, transductions were directly correlated to the therapeutic effect, with low transduction yielding low knockdown and vice versa. Green fluorescent protein (GFP) reporter gene expression was observed with vector incubation times as short as 10 minutes. The highest transductions were seen, after 2-hour 37 degrees C incubation, in UW (62% +/- 6 SEM); they were significantly higher than those in HTK (21% +/- 7 SEM). Neither adenosine nor glutathione, present in UW, provided any increase in transduction when supplemented to HTK, although the addition of hydroxyethyl starch (HES) significantly improved transductions. To rule out size exclusion as a mechanism of HES, IGL-1 was tested but did not result in better transductions than HTK or DMEM. When supplemented to UW, anionic compounds reduced transduction, and this indicated a charge interaction mechanism of HES. In conclusion, this study demonstrates that effective vector delivery can be achieved under conditions of hypothermic liver perfusion. UW provides superior transduction to hepatocytes over nonstarch solutions.
- Subjects :
- Adenosine chemistry
Adenosine pharmacology
Allopurinol chemistry
Allopurinol pharmacology
Cell Line
Glucose chemistry
Glucose pharmacology
Glutathione chemistry
Glutathione pharmacology
Hepatocytes metabolism
Humans
Hypothermia, Induced
Insulin chemistry
Insulin pharmacology
Lentivirus
Liver cytology
Liver metabolism
Mannitol chemistry
Mannitol pharmacology
Organ Preservation
Organ Preservation Solutions chemistry
Perfusion
Potassium Chloride chemistry
Potassium Chloride pharmacology
Procaine chemistry
Procaine pharmacology
Raffinose chemistry
Raffinose pharmacology
Genetic Therapy
Genetic Vectors metabolism
Hepatocytes drug effects
Hydroxyethyl Starch Derivatives chemistry
Liver drug effects
Liver Transplantation
Organ Preservation Solutions pharmacology
Transduction, Genetic
Subjects
Details
- Language :
- English
- ISSN :
- 1527-6473
- Volume :
- 14
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society
- Publication Type :
- Academic Journal
- Accession number :
- 19025921
- Full Text :
- https://doi.org/10.1002/lt.21623