Research Article| Volume 63, ISSUE 2, P388-398, August 2015

Download started.


Pivotal preclinical trial of the spheroid reservoir bioartificial liver

Published:March 26, 2015DOI:

      Background & Aims

      The neuroprotective effect of the spheroid reservoir bioartificial liver (SRBAL) was evaluated in a porcine model of drug-overdose acute liver failure (ALF).


      Healthy pigs were randomized into three groups (standard therapy (ST) alone, ST + No-cell device, ST + SRBAL device) before placement of an implantable intracranial pressure (ICP) monitor and a tunneled central venous catheter. One week later, pigs received bolus infusion of the hepatotoxin D-galactosamine and were followed for up to 90 h.


      At 48 h, all animals had developed encephalopathy and biochemical changes confirming ALF; extracorporeal treatment was initiated and pigs were observed up to 90 h after drug infusion. Pigs treated with the SRBAL, loaded with porcine hepatocyte spheroids, had improved survival (83%, n = 6) compared to ST alone (0%, n = 6, p = 0.003) and No-cell device therapy (17%, n = 6, p = 0.02). Ammonia detoxification, peak levels of serum ammonia and peak ICP, and pig survival were influenced by hepatocyte cell dose, membrane pore size and duration of SRBAL treatment. Hepatocyte spheroids remained highly functional with no decline in mean oxygen consumption from initiation to completion of treatment.


      The SRBAL improved survival in an allogeneic model of drug-overdose ALF. Survival correlated with ammonia detoxification and ICP lowering indicating that hepatocyte spheroids prevented the cerebral manifestations of ALF (brain swelling, herniation, death). Further investigation of SRBAL therapy in a clinical setting is warranted.

      Graphical abstract


      AD (albumin dialysate), ALF (acute liver failure), BAL (bioartificial liver), ECT (extracorporeal therapy), ICP (intracranial pressure), RALF (recovery acute liver failure), ROS (reactive oxygen species), SRBAL (spheroid reservoir bioartificial liver), ST (standard therapy), NC (No-cell)


      Linked Article

      • A new horizon for liver support in acute liver failure
        Journal of HepatologyVol. 63Issue 2
        • Preview
          Acute liver failure (ALF) is a relatively uncommon disease usually affecting young people without preexisting liver disease. Its clinical spectrum includes not only severe derangement of liver function, but also multi-organ failure and a high risk of sepsis, explaining the strong association between ALF and mortality that exceeds 50% [1]. Liver transplantation is the only therapeutic approach that has shown a survival benefit in patients with severe ALF. However, the shortage of organs for transplantation or the existence of contraindications limits the applicability of the procedure.
        • Full-Text
        • PDF
        Open Access
      • Pivotal preclinical trial of the spheroid reservoir bioartificial liver
        Journal of HepatologyVol. 63Issue 4
        • Preview
          We have read with interest the preclinical study of Glorioso et al. [1] on the positive effect of the spheroid reservoir bioartificial liver (SRBAL) treatment in pigs with Acute Liver Failure (ALF) caused by high dose D-galactosamine (RALF study). The presented improvements in survival and histology are very promising and we recognize the challenges (manpower, finances, dealing with variability in pigs) involved with these types of studies.
        • Full-Text
        • PDF
      • Reply to: “Pivotal preclinical trial of the spheroid reservoir bioartificial liver”
        Journal of HepatologyVol. 63Issue 4
        • Preview
          We appreciate the kind words from Dr. Chamuleau and his respected colleagues at Academic Medical Centre, University of Amsterdam. Yes, studies in large animal models of ALF are challenging. Especially recovery studies requiring continuous care of animals over multiple days in succession. As a result, treatment time and membrane porosity were combined to limit our study to three treatment groups (SRBAL device, no cell device, no device) and two device configurations (400 kD membrane porosity, 6 h duration vs.  
        • Full-Text
        • PDF
      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Hepatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Lee W.M.
        • Squires Jr., R.H.
        • Nyberg S.L.
        • Doo E.
        • Hoofnagle J.H.
        Acute liver failure: summary of a workshop.
        Hepatology. 2008; 47: 1401-1415
        • Kjaergard L.
        • Liu J.
        • Als-Nielsen B.
        • et al.
        Artificial and bioartificial support systems for acute and acute-on-chronic liver failure: a systematic review.
        JAMA. 2003; 289: 217-222
        • Demetriou A.A.
        • Brown Jr., R.S.
        • Busuttil R.W.
        • Fair J.
        • McGuire B.M.
        • Rosenthal P.
        • et al.
        Prospective, randomized, multicenter, controlled trial of a bioartificial liver in treating acute liver failure.
        Ann Surg. 2004; 239 (Discussion 667–670): 660-667
        • Saliba F.
        • Camus C.
        • Durand F.
        • Mathurin P.
        • Letierce A.
        • Delafosse B.
        • et al.
        Albumin dialysis with a noncell artificial liver support device in patients with acute liver failure: a randomized controlled trial.
        Ann Intern Med. 2013; 159: 522-531
        • Yu Y.
        • Fisher J.E.
        • Lillegard J.B.
        • Rodysill B.
        • Amiot B.
        • Nyberg S.L.
        Cell therapies for liver diseases.
        Liver Transpl. 2012; 18: 9-21
        • Kelly J.
        • Dalington G.J.
        Modulation of liver specific phenotype in the human hepatoblastoma line Hep G2.
        In Vitro Cell Dev Biol. 1989; 25: 217-222
        • Sauer I.M.
        • Zeilinger K.
        • Obermayer N.
        • Pless G.
        • Grunwald A.
        • Pascher A.
        • et al.
        Primary human liver cells as source for modular extracorporeal liver support – A preliminary report.
        Int J Artif Organs. 2002; 25: 1001-1005
        • Sauer I.
        • Kardassis D.
        • Zeillinger K.
        • Pascher A.
        • Gruenwald A.
        • Pless G.
        • et al.
        Clinical extracorporeal hybrid liver support – Phase 1 study with primary porcine liver cells.
        Xenotransplantation. 2003; 10: 460-469
        • Knowles B.
        • Howe C.
        • Aden D.
        Human hepatocellular carcinoma cell lines secrete the major plasma proteins and Hepatitis B surface antigen.
        Science. 1980; 209: 497-499
        • Nyberg S.L.
        • Remmel R.P.
        • Mann H.J.
        • Peshwa M.V.
        • Hu W.S.
        • Cerra F.B.
        Primary hepatocytes outperform Hep G2 cells as the source of biotransformation functions in a bioartificial liver.
        Ann Surg. 1994; 220: 59-67
        • Mavri-Damelin D.
        • Damelin L.H.
        • Eaton S.
        • Rees M.
        • Selden C.
        • Hodgson H.J.
        Cells for bioartificial liver devices: the human hepatoma-derived cell line C3A produces urea but does not detoxify ammonia.
        Biotechnol Bioeng. 2008; 99: 644-651
        • Mavri-Damelin D.
        • Eaton S.
        • Damelin L.H.
        • Rees M.
        • Hodgson H.J.
        • Selden C.
        Ornithine transcarbamylase and arginase I deficiency are responsible for diminished urea cycle function in the human hepatoblastoma cell line HepG2.
        Int J Biochem Cell Biol. 2007; 39: 555-564
        • Clemmesen J.
        • Larsen F.
        • Kondrug J.
        • Hansen B.
        • Ott P.
        Cerebral herniation in patients with acute liver failure is correlated with arterial ammonia concentration.
        Hepatology. 1999; 29: 648-653
        • Paradis K.
        • Langjord G.
        • Long Z.
        • Heneine W.
        • Sandstrom P.
        • Switzer W.
        • et al.
        Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue.
        Science. 1999; 285: 1236-1241
        • Onions D.E.
        • Witt C.J.
        Xenotransplantation: an overview of microbiological risks and potentials for risk management.
        Rev Sci Tech. 2000; 19: 289-301
        • Nyberg S.L.
        • Hibbs J.R.
        • Hardin J.A.
        • Germer J.J.
        • Persing D.H.
        Transfer of porcine endogenous retrovirus across hollow fiber membranes: significance to a bioartificial liver.
        Transplantation. 1999; 67: 1251-1255
        • Morsiani E.
        • Pazzi P.
        • Puviani A.
        • Brogli M.
        • Valieri L.
        • Gorini P.
        • et al.
        Early experience with a porcine hepatocyte-based bioartificial liver in acute hepatic failure patients.
        Int J Art Organs. 2002; 25: 192-202
        • Nyberg S.L.
        • Yagi T.
        • Matsushita T.
        • Hardin J.
        • Grande J.P.
        • Gibson L.E.
        • et al.
        Membrane barrier of a porcine hepatocyte bioartificial liver.
        Liver Transpl. 2003; 9: 298-305
        • Matsushita T.
        • Amiot B.
        • Hardin J.
        • Platt J.L.
        • Nyberg S.L.
        Membrane pore size impacts performance of a xenogeneic bioartificial liver.
        Transplantation. 2003; 76: 1299-1305
        • Landry J.
        • Bernier D.
        • Ouellet C.
        • Goyette R.
        • Marceau N.
        Spheroidal aggregate culture of rat liver cells: histotypic reorganization, biomatrix deposition, and maintenance of functional activities.
        J Cell Biol. 1985; 101: 914-923
        • Luebke-Wheeler J.
        • Yee L.
        • Nedredal G.
        • Amiot B.
        • Nyberg S.
        E-cadherin protects primary hepatocytes spheroids from cell death by caspase-independent mechanism.
        Cell Transplant. 2010; 18: 1281-1287
        • Abu-Absi S.
        • Friend J.
        • Hansen L.
        • Hu W.-S.
        Structural polarity and functional bile canaliculi in rat hepatocytes spheroids.
        Exp Cell Res. 2002; 274: 56-67
        • Chiarugi P.
        • Giannoni E.
        Anoikis: a necessary death program for anchorage-dependent cells.
        Biochem Pharmacol. 2008; 76: 1352-1364
        • Brophy C.M.
        • Luebke-Wheeler J.L.
        • Amiot B.P.
        • Khan H.
        • Remmel R.P.
        • Rinaldo P.
        • et al.
        Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function.
        Hepatology. 2009; 49: 578-586
        • Nyberg S.L.
        • Hardin J.
        • Amiot B.
        • Argikar U.A.
        • Remmel R.P.
        • Rinaldo P.
        Rapid, large-scale formation of porcine hepatocyte spheroids in a novel spheroid reservoir bioartificial liver.
        Liver Transpl. 2005; 11: 901-910
        • Ho D.
        • Fan S.
        • Woo Y.
        • Zhang Z.
        • Lau C.
        • Wong J.
        Selective plasma filtration for treatment of fulminant hepatic failure induced by D-galactosamine in a pig model.
        Gut. 2002; 50: 869-876
        • Marmarou A.
        • Poll W.
        • Shulman K.
        • Bhagavan H.
        A simple gravimetric technique for measurement of cerebral edema.
        J Neurosurg. 1978; 49: 530-537
        • Sielaff T.D.
        • Hu M.Y.
        • Sridhar R.
        • Groehler K.
        • Olson D.
        • Mann H.J.
        • et al.
        A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture.
        Transplantation. 1995; 59: 1459-1463
        • Lillegard J.B.
        • Fisher J.E.
        • Nedredal G.
        • Luebke-Wheeler J.
        • Bao J.
        • Wang W.
        • et al.
        Normal atmospheric oxygen tension and the use of antioxidants improve hepatocyte spheroid viability and function.
        J Cell Physiol. 2011; 226: 2987-2996
        • Marcus R.
        • Peritz E.
        • Gabriel K.
        On closed testing procedures with special reference to ordered analyses of variance.
        Biometrika. 1976; 63: 655-660
        • Sen S.
        • Rose C.
        • Ytrebo L.
        • Davies N.
        • Nedredal G.
        • Drevland S.
        • et al.
        Effect of albumin dialysis on intracranial pressure in pigs with acute liver failure: a randomized study.
        Crit Care Med. 2006; 34: 158-164
        • Petersen K.
        • Vilstrup H.
        • Tygstrup N.
        Effect of dietary protein on the capacity of urea synthesis in rats.
        Hormone Metab Res. 1990; 22: 612-615
        • Hamberg O.
        • Nielson K.
        • Vilstrup H.
        Effects of increase in protein intake on hepatic efficacy for urea synthesis in healthy subjects and in patients with cirrhosis.
        J Hepatol. 1992; 14: 237-243
        • Kribben A.
        • Gerken G.
        • Haag S.
        • Herget-Rosenthal S.
        • Treichel U.
        • Betz C.
        • et al.
        Effects of Fractionated Plasma Separation and Adsorption on survival in patients with Acute-on-Chronic Liver Failure.
        Gastroenterology. 2012; 142: 782-789
        • Nedredal G.I.
        • Amiot B.P.
        • Nyberg P.
        • Luebke-Wheeler J.
        • Lillegard J.B.
        • McKenzie T.J.
        • et al.
        Optimization of mass transfer for toxin removal and immunoprotection of hepatocytes in a bioartificial liver.
        Biotechnol Bioeng. 2009; 104: 995-1003