By Juliana Puppi, Anil Dhawan (auth.), Anil Dhawan, Robin D. Hughes (eds.)
Mobile treatment utilizing human hepatocytes is being evaluated around the world as a substitute to organ transplantation in sufferers with liver-based metabolic disorder and acute liver failure. the root for medical use has come from the demonstration of efficacy in animal types of acute and protracted liver disease.Protocols were constructed for the isolation of hepatocytes from liver tissue below GMP stipulations and likewise for enhanced tools of cryopreservation, so hepatocytes may be saved for later scientific use. Assays are used to evaluate the standard and serve as of the hepatocytes sooner than transplantation. There are scientific protocols for management of cells at once into the patient’s liver.
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Additional resources for Hepatocyte transplantation: methods and protocols
A1-antitrypsin, a2-macroglobulin). 10% FCS is often used as an addition to WEM for culture of hepatocytes. It can also be used in cryopreservation media but cannot be used for cryopreserving hepatocytes for clinical transplantation. 2. Pre-incubation of hepatocytes with glucose, fructose or a-lipoic acid at 48C prior to cryopreservation has been found to improve thawed hepatocyte viability and function (27). There was no evidence that using the three compounds in combination had an additive effect.
Moscioni, A. , et al. (1995) Repeated intraportal hepatocyte transplantation in analbuminemic rats. Cell Transpl 4, 237–243. 30. De Vree, J. , Bosma, P. , et al. (2000) Correction of liver disease by hepatocyte transplantation in a mouse model of progressive familial intrahepatic cholestasis. Gastroenterology 119, 1720–1730. 31. , Mukhopadhyay, D. , et al. (1998) Long-term, near-total liver replacement by transplantation of isolated hepatocytes in rats treated with retrorsine. Am J Pathol 153, 319–329.
Xenobiotic Metabolism Competence of Hepatocytes The ability of hepatocytes to biotransform xenobiotics is one of the most relevant characteristics of differentiated hepatocytes, and needs to be examined in culture to ensure the appropriate metabolic performance of cells. The 7-ethoxycoumarin O-de-ethylation (ECOD) is catalysed by several CYP isoforms (45), rendering fluorescent 7-hydroxycoumarin that can be easily monitored. The ECOD activity assay can be performed in intact cells and is a good indicator of global P450 activities.
Hepatocyte transplantation: methods and protocols by Juliana Puppi, Anil Dhawan (auth.), Anil Dhawan, Robin D. Hughes (eds.)