Hybrid artificial liver using hepatocyte organoid culture.

We developed 2 types of hybrid artificial liver modules using hepatocyte organoid culture. One was a polyurethane foam (PUF)/hepatocyte spheroid packed-bed module. Hepatocytes spontaneously formed spheroids in the PUF pores, and they maintained liver-specific functions well for at least 2 weeks in vitro. As a preclinical experiment, a hybrid artificial liver with 200 g porcine hepatocytes was applied to a pig (25 kg) with liver failure and showed that the hybrid artificial liver was effective in support of liver functions and stabilization of general conditions. We established a new technique of hepatocyte organoid formation using centrifugal force. A hepatocyte organoid formed by centrifugation in hollow fibers maintained functions for more than 4 months in vitro. We developed a new sinusoid-like structure module having hollow fibers arranged by spacers in a micro-regular arrangement. Inoculated hepatocytes in the extra-fiber space of the module formed the organoid by centrifugation, and they maintained the functions for at least 1 month in vitro. The results indicated that this module seems to be promising as a hybrid artificial liver.

[1]  M. Suzuki,et al.  Large-Scale Preparation and Function of Porcine Hepatocyte Spheroids , 1996, The International journal of artificial organs.

[2]  K. Nakazawa,et al.  Intensive promotion of spheroid formation by soluble factors in a hepatocyte-conditioned medium , 2000, Journal of biomaterials science. Polymer edition.

[3]  S. Ash,et al.  Cytokines and endotoxin removal by sorbents and its application in push-pull sorbent-based pheresis: the BioLogic-DTPF System. , 1999, Artificial organs.

[4]  A. Demetriou,et al.  Clinical experience with a bioartificial liver in the treatment of severe liver failure. A phase I clinical trial. , 1997, Annals of surgery.

[5]  K. Sugimachi,et al.  Development of hybrid artificial liver support system using spheroid culture and application to warm ischemic liver failure in dog and pig as a preclinical test , 1998 .

[6]  P Neuhaus,et al.  Improved hepatocyte in vitro maintenance in a culture model with woven multicompartment capillary systems: Electron microscopy studies , 1995, Hepatology.

[7]  P. Sawyer,et al.  The development of polymeric cardiovascular collagen prostheses. , 2008, Artificial organs.

[8]  Yasuhiro Takeuchi,et al.  Infection of human cells by an endogenous retrovirus of pigs , 1997, Nature Medicine.

[9]  K. Asano,et al.  Continued high albumin production by multicellular spheroids of adult rat hepatocytes formed in the presence of liver-derived proteoglycans. , 1989, Biochemical and biophysical research communications.

[10]  T. Giorgio,et al.  Development of a bioartificial liver: Properties and function of a hollow‐fiber module inoculated with liver cells , 1993, Hepatology.

[11]  S. Matsubara,et al.  Combination of plasma exchange and continuous hemofiltration as temporary metabolic support for patients with acute liver failure. , 1994, Artificial organs.

[12]  K Sugimachi,et al.  Modulation of immunologic reactions between cultured porcine hepatocytes and human sera. , 1999, ASAIO journal.

[13]  K Sugimachi,et al.  Evaluation of a hybrid artificial liver using a polyurethane foam packed-Bed culture system in dogs. , 1999, The Journal of surgical research.

[14]  Kohji Nakazawa,et al.  Hepatocyte Spheroids in Polyurethane Foams: Functional Analysis and Application for a Hybrid Artificial Liver , 1998 .

[15]  J. Chowdhury,et al.  Characterization and Evaluation of Detoxification Functions of a Nontumorigenic Immortalized Porcine Hepatocyte Cell Line (HepLiu1) , 1999, Cell transplantation.

[16]  V. Dixit,et al.  Artificial liver support: state of the art. , 1996, Scandinavian journal of gastroenterology. Supplement.

[17]  W. Heneine,et al.  Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue. The XEN 111 Study Group. , 1999, Science.

[18]  N. Sussman,et al.  Reversal of fulminant hepatic failure using an extracorporeal liver assist device , 1992, Hepatology.

[19]  F. Alvarez,et al.  Long-term culture of adult rat hepatocyte spheroids. , 1992, Experimental cell research.

[20]  K. Oldhafer,et al.  A novel bioreactor design for in vitro reconstruction of in vivo liver characteristics. , 1995, Artificial organs.

[21]  K. Sugimachi,et al.  Development of a Hybrid Artificial Liver using a Polyurethane Foam/Hepatocyte-Spheroid Packed-Bed Module , 2000, The International journal of artificial organs.

[22]  K. Yoshizato,et al.  Morphological and immuno-cytochemical characterization of a hetero-spheroid composed of fibroblasts and hepatocytes. , 1992, Journal of cell science.

[23]  L. Flendrig,et al.  Semipermeable hollow fiber membranes in hepatocyte bioreactors: a prerequisite for a successful bioartificial liver? , 2008, Artificial organs.

[24]  C. Mullon,et al.  Evidence of absence of porcine endogenous retrovirus (PERV) infection in patients treated with a bioartificial liver support system. , 1999, Artificial organs.

[25]  N. Sussman,et al.  The Hepatix extracorporeal liver assist device: initial clinical experience. , 1994, Artificial organs.

[26]  H. Mizumoto,et al.  Formation of a spherical multicellular aggregate (spheroid) of animal cells in the pores of polyurethane foam as a cell culture substratum and its application to a hybrid artificial liver. , 1998, Journal of biomaterials science. Polymer edition.

[27]  K. Sugimachi,et al.  Conditions Required for a Hybrid Artificial Liver Support System using a PUF/Hepatocyte-Spheroid Packed-Bed Module and it's use in Dogs with Liver Failure , 2000, The International journal of artificial organs.

[28]  Y. Sakai,et al.  Comparisons of Porcine Hepatocyte Spheroids and Single Hepatocytes in the Non-Woven Fabric Bioartificial Liver Module , 1996, The International journal of artificial organs.

[29]  M. Löhr,et al.  Molecular adsorbent recycling system (MARS): clinical results of a new membrane-based blood purification system for bioartificial liver support. , 1999, Artificial organs.

[30]  S. Ash.  Hemodiabsorption in treatment of acute hepatic failure and chronic cirrhosis with ascites. , 1994, Artificial organs.