title: Evaluation of encapsulated liver cell spheroids in a fluidised-bed bioartificial liver for treatment of ischaemic acute liver failure in pigs in a translational setting.
abstract: Liver failure is an increasing problem. Donor-organ shortage results in patients dying before receiving a transplant. Since the liver can regenerate, alternative therapies providing temporary liver-support are sought. A bioartificial-liver would temporarily substitute function in liver failure buying time for liver regeneration/organ-procurement. Our aim: to develop a prototype bioartificial-liver-machine (BAL) comprising a human liver-derived cell-line, cultured to phenotypic competence and deliverable in a clinical setting to sites distant from its preparation. The objective of this study was to determine whether its use would improve functional parameters of liver failure in pigs with acute liver failure, to provide proof-of-principle. HepG2 cells encapsulated in alginate-beads, proliferated in a fluidised-bed-bioreactor providing a biomass of 4-6 × 10(10)cells, were transported from preparation-laboratory to point-of-use operating theatre (6000 miles) under perfluorodecalin at ambient temperature. Irreversible ischaemic liver failure was induced in anaesthetised pigs, after portal-systemic-shunt, by hepatic-artery-ligation. Biochemical parameters, intracranial pressure, and functional-clotting were measured in animals connected in an extracorporeal bioartificial-liver circuit. Efficacy was demonstrated comparing outcomes between animals connected to a circuit containing alginate-encapsulated cells (Cell-bead BAL), and those connected to circuit containing alginate capsules without cells (Empty-bead BAL). Cells of the biomass met regulatory standards for sterility and provenance. All animals developed progressive liver-failure after ischaemia induction. Efficacy of BAL was demonstrated since animals connected to a functional biomass (+ cells) had significantly smaller rises in intracranial pressure, lower ammonia levels, more bilirubin conjugation, improved acidosis and clotting restoration compared to animals connected to the circuit without cells. In the +cell group, human proteins accumulated in pigs' plasma. Delivery of biomass using a short-term cold-chain enabled transport and use without loss of function over 3 days. Thus, a fluidised-bed bioreactor containing alginate-encapsulated HepG2 cell-spheroids improved important parameters of acute liver failure in pigs. The system can readily be up-scaled and transported to point-of-use justifying development at clinical scale.
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Clare
authors.initials: C
authors.lastname: Selden
authors.affiliation: Division of Hepatology, Department of Medicine, University of Cape Town, Groote Schuur Hospital, Cape Town, Western Cape, South Africa.
authors.forename: Catherine Wendy
authors.initials: CW
authors.lastname: Spearman
authors.affiliation: Department of Surgery, University of Cape Town, Groote Schuur Hospital, Cape Town, Western Cape, South Africa.
authors.forename: Delawir
authors.initials: D
authors.lastname: Kahn
authors.affiliation: Department of Anaesthetics, University of Cape Town, Groote Schuur Hospital, Cape Town, Western Cape, South Africa.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Miller
authors.affiliation: Department Neurosurgery, Red Cross Children's Hospital, University of Cape Town, Cape Town, Western Cape, South Africa.
authors.forename: Anthony
authors.initials: A
authors.lastname: Figaji
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Eloy
authors.initials: E
authors.lastname: Erro
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: James
authors.initials: J
authors.lastname: Bundy
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Isobel
authors.initials: I
authors.lastname: Massie
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Sherri-Ann
authors.initials: SA
authors.lastname: Chalmers
authors.affiliation: Department of Surgery, University of Cape Town, Groote Schuur Hospital, Cape Town, Western Cape, South Africa.
authors.forename: Hiram
authors.initials: H
authors.lastname: Arendse
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Aude
authors.initials: A
authors.lastname: Gautier
authors.affiliation: Biochemistry Department, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Sharratt
authors.affiliation: Division of Surgery and Interventional Science, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Barry
authors.initials: B
authors.lastname: Fuller
authors.affiliation: University College London Institute for Liver & Digestive Health, University College London Medical School, Royal Free Hospital Campus, Hampstead, London, United Kingdom.
authors.forename: Humphrey
authors.initials: H
authors.lastname: Hodgson
mesh_terms: D000818:Animals; D019149:Bioreactors; D002470:Cell Survival / Q000502:physiology; D005260:Female; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D017114:Liver Failure, Acute / Q000378:metabolism / Q000473:pathology* / Q000601:surgery*; D019164:Liver, Artificial; D018874:Spheroids, Cellular / Q000166:cytology* / Q000378:metabolism; D013552:Swine