Factors affecting hepatocyte isolation, engraftment, and replication in an in vivo model

Abstract

Human hepatocyte transplantation is an alternative treatment for acute liver failure and liver diseases involving enzyme deficiencies. Although it has been successfully applied in selected recipients, both isolation and transplantation outcomes have the potential to be improved by better donor selection. This study assessed the impact of various donor variables on isolation outcomes (yield and viability) and posttransplant engraftment, using the SCID/Alb-uPA (severe combined immunodeficient/urokinase type plasminogen activator under the control of an albumin promoter) human liver chimeric mouse model. Human hepatocytes were obtained from 90 human liver donor specimens and were transplanted into 3942 mice. Multivariate analysis revealed improved viability with younger donors (P ¼ 0.038) as well as with shorter warm ischemic time (P ¼ 0.012). Hepatocyte engraftment, assessed by the posttransplant level of serum human a1-antitrypsin, was improved with shorter warm ischemia time. Hepatocytes isolated from older donors (!60 years) had lower viability and posttransplant engraftment (P 0.01). In conclusion, the selection of young donors (<60 years) and rapid liver specimen retrieval, allowing for shorter warm ischemia time, are key determinants for the success of both the isolation of high viability human hepatocytes and their subsequent posttransplantation capacity for engraftment and expansion. Liver Transpl 16:974-982,

Recapitulation of treatment response patterns in a novel humanized mouse model for chronic hepatitis B virus infection

“…A variety of liver injury models, including Alb-uPA (Dandri et al, 2001; Mercer et al, 2001; Meuleman et al, 2005), MUP-uPA (Carpentier et al, 2014; Heo et al, 2006), Alb-HSV-tk (Hasegawa et al, 2011) and FAH −/− mice (Azuma et al, 2007; Bissig et al, 2007; He et al, 2010), are being used to facilitate human hepatic engraftment on immunodeficient mouse backgrounds. Engraftment levels vary due to a number of factors, including the specific liver injury, the severity of the immunodeficiency and the quality (donor age, underlying liver diseases, fresh or cryopreserved) of the human hepatocytes (Kawahara et al, 2010; Vanwolleghem et al, 2010). FAH deficient mice are particularly attractive as they can be easily propagated, and the liver injury can be simply controlled by addition or removal of the liver protective drug NTBC.…”

Advances in experimental systems to study hepatitis C virus in vitro and in vivo

“…The advantage of using inducible liver injury models over uPAexpressing mice is that the murine liver injury is reversible, thus enabling longer life spans of reconstituted animals by cycling liver injury. Reconstitution levels with human hepatocytes can be conveniently monitored using human-specific serum albumin Elisa assays and studies revealed that 1 mg/mL of human albumin, which is the minimum level for infection studies with HCV correlates to 10% of the liver being engrafted with human hepatocytes (Kawahara et al, 2010). Strikingly, not only HCVcc can be used to infect human liver-chimeric mice but also patient isolates of different genotypes have successfully be used to launch infection (Mercer et al, 2001).…”

Visualizing hepatitis C virus infection in humanized mice

“…Resulting human liver chimeric Alb-uPA (Mercer et al, 2001; Meuleman et al, 2005), MUP-uPA (Tesfaye et al, 2013), HSV-tk (Kosaka et al, 2013) and FAH-/- (Bissig et al, 2010) mice are used for infection with cell culture derived HCV and patient isolates. Susceptibility to HCV infection correlates with a high human hepatic chimerism (Bissig et al, 2010; Kawahara et al, 2010; Vanwolleghem et al, 2010). …”