Stand der Hepatozytentransplantation: Risiko oder Chance?

 

 Buy Article Permissions and Reprints

Zusammenfassung

Bei unterschiedlichen Lebererkrankungen wird seit einigen Jahren alternativ zur orthotopen Lebertransplantation die Hepatozytentransplantation als neuer experimenteller Therapieansatz untersucht. Neben dem kurativen Ansatz bei genetischen Stoffwechseldefekten (Hämophilie, familiäre Hypercholesterinämie, etc.) ist die Hepatozytentransplantation als Ü berbrückung der Wartezeit auf ein geeignetes Lebertransplantat angedacht. Präklinische Studien am Tiermodell konnten diesbezüglich zeigen, dass sich intraperitoneal, intraportal oder über die Empfängermilz infundierte Hepatozyten in der erkrankten Leber absiedeln. Dies ermöglicht sowohl die Korrektur verschiedener angeborener Stoffwechselerkrankungen als auch eine verbesserte Überlebensrate beim akuten Leberversagen. Erstmalig wurde 1992 in einer klinischen Studie an 10 Patienten die Transplantation isolierter Hepatozyten vorgenommen. Fox u. Mitarb. beschrieben 1998 die erfolgreiche Transplantation allogener Leberzellen bei einem Kind mit Crigler-Najjar-Syndrom. Im gleichen Zeitraum durchgeführte Studien von Strom et al. bzw. Bilir et al. belegen die erfolgreiche Leberzelltransplantation zur transienten Behandlung des akuten Leberversagens. Grundsätzliche Voraussetzung für die Durchführung der Hepatozytentransplantation ist eine ausreichende Zellzahl differenzierter Hepatozyten. Um den Einsatz isolierter humaner Leberzellen in der Klinik ermöglichen zu können, ist die Entwicklung einer In-vitro- Kulturtechnik erforderlich, die sowohl eine reproduzierbare und hinreichend genügende Zellteilungsrate in Hepatozyten zu induzieren vermag als auch den Erhalt der physiologischen Funktion der Zellen garantiert. In der Arbeit wird eine Übersicht über die verschiedenen Ansätze der Kultivierung humaner Hepatozyten gegeben sowie auf alternative Zellquellen (tierische Hepatozyten, immortalisierte Zelllinien humanen Ursprungs, fetale Zellen bzw. Stammzellen), die zur Hepatozytentransplantation verwendet werden könnten, eingegangen und diese evaluiert.

Abstract

Over the past few years, hepatocyte transplantation has been considered as an alternative method for orthotopic liver transplantation for the treatment of various liver diseases. Beside curative approach for genetic metabolic deficiences (familial hypercholesterolemia, hemophilia, etc.), it could be a useful tool for bridging the waiting period until an appropriate donor organ is obtained. In preclinical animal studies, hepatocytes injected intraperitoneally, intraportally or into the spleen settle down in the diseased liver. This enables genetic modification to correct inborn metabolic deficiences and improves survival in acute liver failure. In 1992, the first clinical transplantation of isolated hepatocytes in 10 patients was performed. In 1998, Fox and coworkers described the succesful transplantation of allogeneic liver cells in a child with Crigler-Najjar syndrome. Accomplished studies of Strom et al. resp. Bilir et al. of the same year proved the effectiveness of liver cell transplantation for transient treatment of acute liver failure. Prerequisite of this cell-based therapeutic strategy is a sufficient amount of highly differentiated hepatocytes, hence, a well established in-vitro cell-culture technique is necessary to yield a reproducible number of proliferating hepatocytes and to preserve the physiological cell function. This review discusses the different experimental approaches regarding the cultivation of human hepatocytes and also the use of alternative cell sources (like animal hepatocytes, immortalized cells of human origin, progenitor cells from fetal human liver/liver stem cells) for hepatocyte transplantation.

Literatur

• 1 Auth M K, Keitzer R A, Scholz M, Blaheta R A, Hottenrott E C, Herrmann G, Encke A, Markus B H. Establishment and immunological characterization of cultured human gallbladder epithelial cells.  Hepatology. 1993;  18 546-555
  CrossrefPubMedGoogle Scholar
• 2 Baccarani U, Donini A, Risaliti A, Piccolo G, Dialti V, Cautero N, Degrassi A, Sirchia G, Bresadola F. Steatotic versus cirrhotic livers as a source for human hepatocyte isolation.  Transplant Proc. 2001;  33 664-665
  CrossrefPubMedGoogle Scholar
• 3 Bader A, Knop E, Boker K, Fruhauf N, Schuttler W, Oldhafer K, Burkhard R, Pichlmaayr R, Sewing K F. A novel bioreactor design for in vitro reconstruction of in vivo liver characteristics.  Artif Organs. 1995;  19 368-374
  PubMedGoogle Scholar
• 4 Benedetti E, Kirby J P, Asolati M, Blanchard J, Ward M G, Williams R, Hewett T A, Fontaine M, Pollak R. Intrasplenic hepatocyte allotransplantation in dalmatian dog with and without cyclosporine immunosuppression.  Transplantation. 1997;  63 1206-1209
  PubMedGoogle Scholar
• 5 Berthiaume F, Moghe P V, Toner M, Yarmush M L. Effect of extracellular matrix topology on cell structure, function, and physiological responsivness: hepatocytes cultured in a sandwich configuration.  FASEB J. 1996;  10 1471-1484
  PubMedGoogle Scholar
• 6 Bhatia S N, Balis U J, Yarmush M L, Toner M. Effect of cell-cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells.  FASEB J. 1999;  13 1883-1900
  PubMedGoogle Scholar
• 7 Bilir B M, Guinette D, Karrer F, Kumpe D A, Krysl J, Stephens J, McGavran L, Ostrowska A, Durham J. Hepatocyte transplantation in acute liver failure.  Liver Transpl. 2000;  6 32-40
  PubMedGoogle Scholar
• 8 Block G D, Locker J, Bowen W C, Peterson B E, Katyal S, Strom S C, Riley T, Howard T A, Michalopoulos G K. Population expansion, clonal growth, and specific differentiation patterns in primary cultures of hepatocytes induced by HGF/SF, EGF and TGF alpha in a chemically defined (HGM) medium.  J Cell Biol. 1996;  132 1133-1149
  CrossrefPubMedGoogle Scholar
• 9 Bruni S, Chang T M. Kinetic analysis of UDP-glucuronosyltransferase in bilirubin conjugation by encapsulated hepatocytes for transplantation in Gunn rats.  Artif Organs. 1995;  19 449-457
  PubMedGoogle Scholar
• 10 Busse B, Gerlach J C. Bioreactors for hybrid liver support: histological aspects and novel designs.  Ann N Y Acad Sci. 1999;  875 326-339
  PubMedGoogle Scholar
• 11 Busse B, Smith M D, Gerlach J C. Treatment of acute liver failure: hybrid liver support. A critical overview.  Langenbecks Arch Surg. 1999;  384 588-599
  CrossrefPubMedGoogle Scholar
• 12 Clement B, Guguen-Guillouzo C, Grimaud J, Rissel M, Guillouzo A. Effect of hydrocortisone on deposition of types I and IV collagen in primary culture of rat hepatocytes.  Cell Mol Biol. 1988;  34 449-460
  PubMedGoogle Scholar
• 13 Dunn T B, Kumins N H, Raofi V, Holman D M, Mikalov M, Blanchard J, Law W R, Rastellini C, Benedetti E. Multiple intrasplenic hepatocyte transplantation in the dalmatian dog.  Surgery. 2000;  127 193-199
  CrossrefPubMedGoogle Scholar
• 14 Eguchi S, Rozga J, Lebow L T, Chen S C, Wang C C, Rosenthal R, Fogli L, Hewitt W R, Middleton Y, Demetriou A A. Treatment of hypercholesterolemia in the Watanabe rabbit using allogeneic hepatocellular transplantation under a regeneration stimulus.  Transplantation. 1996;  62 588-593
  PubMedGoogle Scholar
• 15 Eurotransplant International Foundation .Annual Report. 2000
  Google Scholar
• 16 Fisher R A, Bu D, Thompson M, Tisnado J, Prasad U, Sterling R, Posner M, Strom S. Defining hepatocellular chimerism in a liver failure patient bridged with hepatocyte infusion.  Transplantation. 2000;  69 303-307
  PubMedGoogle Scholar
• 17 Fox I J, Chowdhury J R, Kaufman S S, Goertzen T C, Chowdhury N I, Warkentin P I, Dorko K, Sauter B V, Strom S C. Treatment of the Crigler-Najjar Syndrome Type I with hepatocyte transplantation.  N Engl J Med. 1998;  338 1422-1426
  PubMedGoogle Scholar
• 18 Gagandeep S, Rajvanshi P, Sokhi R P, Slehria S, Palestro C J, Bhargava K K, Gupta S. Transplanted hepatocytes engraft, survive, and proliferate in the liver of rats with carbon tetrachloride-induced cirrhosis.  J Pathol. 2000;  191 78-85
  PubMedGoogle Scholar
• 19 Gagandeep S, Sokhi R, Slehria S, Gorla G R, Furgiuele J, DePinho R A, Gupta S. Hepatocyte transplantation improves survival in mice with liver toxicity induced by hepatic overexpression of Mad1 transcription factor.  Mol Ther. 2000;  1 358-365
  CrossrefPubMedGoogle Scholar
• 20 Gerlach J C. Development of a hybrid liver support system: a review.  Int J Artif Organs. 1996;  19 645-654
  PubMedGoogle Scholar
• 21 Gottlieb D I, Huettner J E. An in vitro pathway from embryonic stem cells to neurons and glia.  Cells Tissues Organs. 1999;  165 165-172
  CrossrefPubMedGoogle Scholar
• 22 Gregory P G, Conolly C K, Toner M, Sullivan S J. In vitro characterization of porcine hepatocyte function.  Cell Transplant. 2000;  9 1-10
  PubMedGoogle Scholar
• 23 Grompe M. Therapeutic liver repopulation for the treatment of metabolic liver diseases.  Hum Cell. 1999;  12 171-180
  PubMedGoogle Scholar
• 24 Guha C, Parashar B, Deb N J, Sharma A, Gorla G R, Alfieri A, Roy-Chowdhury N, Roy-Chowdhury J, Vikram B. Liver irradiation: a potential preparative regimen for hepatocyte transplantation.  Int J Radiat Oncol Biol Phys. 2001;  49 451-457
  CrossrefPubMedGoogle Scholar
• 25 Harper A M, McBride M A, Ellison E D. The UNOS OPTN waiting list, 1988 - 1998. Clin Transpl 1999; 71 - 82
  Google Scholar
• 26 Hescheler J, Fleischmann B K, Wartenberg M, Bloch W, Kolossov E, Ji G, Addicks K, Sauer H. Establishment of ionic channels and signalling cascades in the embryonic stem cell-derived primitive endoderm and cardiovascular system.  Cells Tissues Organs. 1999;  165 153-164
  CrossrefPubMedGoogle Scholar
• 27 Hino O, Kitagawa T, Nomura K, Ohtake K, Cui L, Furuta Y, Aizawa S. Hepatocarcinogenesis in transgenic mice carrying albumin-promoted SV40 T antigen gene.  Jpn J Cancer Res. 1991;  82 1226-1233
  PubMedGoogle Scholar
• 28 Hodgkinson C P, Wright M C, Paine A J. Fibronectin-mediated hepatocyte shape change reprograms cytochrome P450 2C11 gene expression via an integrin-signaled induction of ribunuclease activity.  Mol Pharmacol. 2000;  58 976-981
  PubMedGoogle Scholar
• 29 Hole N. Embryonic stem cell-derived haematopoiesis.  Cells Tissues Organs. 1999;  165 181-189
  CrossrefPubMedGoogle Scholar
• 30 Ikebukuro H, Inagaki M, Mito M, Kasai S, Ogawa K, Nozawa M. Prolonged function of hepatocytes transplantated into the spleens of Ngase analbumineic rats.  Eur Surg Res. 1999;  31 39-47
  PubMedGoogle Scholar
• 31 Kamohara Y, Rozga J, Demetriou A A. Artifical liver: review and Cedars-Sinai experience.  J Hepatobiliary Pancreat Surg. 1998;  5 273-285
  CrossrefPubMedGoogle Scholar
• 32 Keeffe E B. Liver transplantation current status and novel approaches to liver replacement.  Gastroenterology. 2001;  120 749-762
  PubMedGoogle Scholar
• 33 Kim W H, Lee J H, Han S U, Jin Y M, Kwak Y S, Wang H J, Kim M W. Systematic analysis of the effects of hepatocyte transplantation on rats with acute liver failure.  Hepatogastroenterology. 2000;  47 371-374
  PubMedGoogle Scholar
• 34 Kobayashi N, Noguchi H, Fujiwara T, Tanaka N. Xenotransplantation of immotalized human hepatocytes for experimental acute liver failure in rats.  Transplant Proc. 2000;  32 1123-1124
  CrossrefPubMedGoogle Scholar
• 35 Kocken J M, Borel Rinkes I H, Bijma A M, de Roos W K, Bowman E, Terpstra O T, Sinaasappel M. Correction of an inborn error of metabolism by intraportal hepatocyte transplantation in a dog model.  Transplantation. 1996;  62 358-364
  PubMedGoogle Scholar
• 36 Kono Y, Yang S, Roberts E A. Extendet primary culture of human hepatocytes in a collagen gel sandwich system.  In Vitro Cell Dev Biol Anim. 1997;  33 467-472
  PubMedGoogle Scholar
• 37 Lagasse E, Connors H, Al-Dhalimy M, Reitsma M, Dohse M, Osborne L, Wang X, Finegold M, Weissman I L, Grompe M. Purified hematopoietic stem cells can differentiate into hepatocytes in vivo.  Nat Med. 2000;  6 1229-1234
  CrossrefPubMedGoogle Scholar
• 38 Leckel K, Blaheta R A, Boost K A, Kim H J, Markus B H. In-vitro Analysen zum klinischen Einsatz isolierter humaner Hepatozyten. Chirurgisches Forum 2001 für experimentelle und klinische Forschung.  Forumband. 2001;  30 281-283
  PubMedGoogle Scholar
• 39 Leckel K, Blaheta R A, Boost K A, Krätschell R, Auth M K, Markus B H. Development of a cell culture modell to allow controlled modification of the differentiation status of human hepatocytes. Zur Publikation eingereicht in J Hepatol
  Google Scholar
• 40 Lee J, Morgan J R, Tompkins R G, Jarmush M L. Proline-mediated enhancement of hepatocyte function in a collagen sandwich culture configuration.  FASEB L. 1993;  7 586-591
  PubMedGoogle Scholar
• 41 LeTissier P, Stoye J P, Takeuchi Y, Patience C, Weiss R A. Two sets of human-topic pig retrovirus.  Nature. 1997;  389 681-682
  CrossrefPubMedGoogle Scholar
• 42 Malhi H, Gupta S. Hepatocyte transplantation: new horizons and challenges.  J Hepatobiliary Pancreat Surg. 2001;  8 40-50
  CrossrefPubMedGoogle Scholar
• 43 Markus P M, Becker H. Hepatocelluläre Transplantation - von den Anfängen bis zum klinischen Einsatz.  Chirurg. 1999;  70 162-168
  PubMedGoogle Scholar
• 44 Michalopoulos G K, Bowen W, Nussler A K, Becich M J, Howard T A. Comparative analysis of mitogenic and morphogenic effects of HGF and EGF on rat and human hepatocytes maintained in collagen gels.  J Cell Physiol. 1993;  156 443-452
  CrossrefPubMedGoogle Scholar
• 45 Mitchell C, Mignon A, Guidotti J E, Besnard S, Fabre M, Duverger N, Parlier D, Tedgui A, Kahn A, Gilgenkrantz H. Therapeutic liver repopulation in a mouse model of hypercholesterolemia.  Hum Mol Genet. 2000;  9 1597-1602
  CrossrefPubMedGoogle Scholar
• 46 Mito M, Kusano M, Kawaura Y. Hepatocyte transplantation in man.  Transplant Proc. 1992;  24 3052-3053
  PubMedGoogle Scholar
• 47 Naik S, Trenkler D, Santangini H, Pan J, Jaureguitto H O. Isolation and culture of porcine hepatocytes for artificial liver support.  Cell Transplant. 1996;  5 107-115
  PubMedGoogle Scholar
• 48 Nakamura J, Okamoto T, Schumacher I K, Tabei I, Chowdhury N R, Chowdhury J R, Fox I J. Treatment of surgically induced acute liver failure by transplantation of conditionally immortalised hepatocytes.  Transplantation. 1997;  63 1541-1547
  PubMedGoogle Scholar
• 49 Nozawa K, Kurumiya Y, Yamamoto A, Isobe Y, Suzuki M, Yoshida S. Up-regulation of telomerase in primary cultured rat hepatocytes.  J Biochem. 1999;  126 361-367
  PubMedGoogle Scholar
• 50 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 a source of biotransformation functions in a bioartifical liver.  Ann Surg. 1994;  220 59-67
  PubMedGoogle Scholar
• 51 Oren R, Dabeva M D, Petkov P M, Hurston E, Laconi E, Shafritz D A. Restoration of serum albumin levels in nagase analbumineic rats by hepatocyte transplantation.  Hepatology. 1999;  29 75-81
  CrossrefPubMedGoogle Scholar
• 52 Osawa M, Hanada K, Hamada H, Nakauchi H. Long-tern lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell.  Science. 1996;  273 242-245
  PubMedGoogle Scholar
• 53 Ott M, Schmidt H H, Cichon G, Manns M P. Emerging therapies in hepatology: liver-directed gene transfer and hepatocyte transplantation.  Cells Tissues Organs. 2000;  167 8187
  PubMedGoogle Scholar
• 54 Patience C, Takeuchi Y, Weiss R A. Infection of human cells by an endogenous retrovirus of pigs.  Nat Med. 1997;  54 282-286
  PubMedGoogle Scholar
• 55 Petersen B E, Bowen W C, Patrene K D, Mars W M, Sullivan A K, Murase N, Boggs S S, Greenberger J S, Goff J P. Bone marrow as a potential source of hepatic oval cells.  Science. 1999;  284 1168-1170
  CrossrefPubMedGoogle Scholar
• 56 Puviani A C, Ottolenghi C, Tassinari B, Pazzi P, Marsiani E. An update on high-yield hepatocyte isolation methods and on the potential clinical use of isolated liver cells.  Comp Biochem Physiol. 1998;  121 99-109
  PubMedGoogle Scholar
• 57 Ransford R, Gunson B, Mayer D, Neuberger J, Christensen E. Effect on outcome of the lengthening waiting list for liver transplantation.  Gut. 2000;  47 441-443
  CrossrefPubMedGoogle Scholar
• 58 Raper S E, Grossmann M, Rader D J, Thoene J G, Clark BJ 3  rd , Kolansky D M, Muller D W, Wilson J M. Safety and feasibility of liver directed gene therapy for homozygous familial hypercholesterolemia.  Ann Surg. 1996;  223 116-126
  CrossrefPubMedGoogle Scholar
• 59 Reyes J, Rubenstein W S, Mieles L. The use of cultured hepatocyte infusion via the portal vein for the treatment of ornithine transcarbamylase deficiency by transplantation of enzymatically competent ABO/Rh-matched cells.  Hepatology. 1996;  24 308-313
  PubMedGoogle Scholar
• 60 Riordan S M, Williams R. Acute liver failure: targeted artificial and hepatocyte-based support of liver regeneration and reversal mutiorgan failure.  J Hepatol. 2000;  32 63-76
  PubMedGoogle Scholar
• 61 Riordan S M, Williams R. Extracorporal support and hepatocyte transplantation in acute liver failure and cirrhosis.  J Gastroenterol Hepatol. 1999;  14 757-770
  CrossrefPubMedGoogle Scholar
• 62 Rodes J, Navasa M. Liver transplantation and quality of live.  Can J Gatroenterol. 2000;  14 693-699
  PubMedGoogle Scholar
• 63 Rozga J, Morsiani E, LePage A D, Morsiani E, Moscioni A D, Hoffman A, Sher L, Villamil F, Woolf G, McGrath M. A bioartificial liver to treat severe acute liver failure.  Ann Surg. 1994;  219 538-544
  PubMedGoogle Scholar
• 64 Schroder A J, Blaheta R A, Scholz M, Encke A, Markus B H. Isolation and separation of human adult hepatocytes from resected liver: cellular yield and purity using different methods.  Zentralbl Chir. 1994;  119 127-138
  PubMedGoogle Scholar
• 65 Schroder A J, Blaheta R A, Scholz M, Kronenberger B, Encke A, Markus B H. Effects of proinflammatory cytokines on cultivated primary human hepatocytes. Fluorometric measurement of intercellular adhesion molecule-1 and human leukocyte antigen -A, -B, -C, and -DR expression.  Transplantation. 1995;  59 1023-1028
  PubMedGoogle Scholar
• 66 Sierra E, Maganto P, Codesal J, Mula N, Cubero J, Arza E, Castillo-Olivares J L, Arahuetes R M. Liver gene expression and increase in albumin synthesis by fetal hepatocytes transplanted into analbuminemic rats.  Life Sci. 2000;  67 2417-2432
  CrossrefPubMedGoogle Scholar
• 67 Stange J, Mitzner S. Cell sources for bioartifical liver support.  Int J Artif Organs. 1996;  19 14-17
  PubMedGoogle Scholar
• 68 Stevens L C. The origin and development of testicular, ovarian, and embryo-derived teratomas. In: Silver LM, Martin GR, Strickland S (Hrsg). Teratocarcinoma Stem Cells. Cold Spring Harbor Laboratora Press, Cold Spring Harbor, New York 1983; 23 - 36
  Google Scholar
• 69 Strom S C, Chowdhury J R, Fox I J. Hepatocyte transplantation for the treatment of human disease.  Semin Liver Dis. 1999;  19 39-48
  PubMedGoogle Scholar
• 70 Strom S C, Fisher R A, Thompson M T, Sanyal A J, Cole P E, Ham J M, Posner M P. Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure.  Transplantation. 1997;  63 559-569
  PubMedGoogle Scholar
• 71 Sundback C A, Vacanti J P. Alternatives to liver transplantation: from hepatocyte transplantation to tissue-engineered organs.  Gastroenterology. 2000;  118 438-442
  PubMedGoogle Scholar
• 72 Suc B, Chalem Y, Golmard J L. Analysis of the French liver transplant waiting list, 1992 - 1996.  Transplantation. 2000;  69 515-522
  PubMedGoogle Scholar
• 73 Sugiyama N, Eguchi S, Kawazoe Y, Miyamoto S, Kamohara Y, Fujioka H, Furui J, Sato M, Ishii T, Kanematsu T. Intraportal administration of low-dose recombinant human hepatocyte growth factor anhances effects of hepatocellular transplantation.  Hepatogastroenterology. 2000;  47 1245-1249
  PubMedGoogle Scholar
• 74 Sutherland D E, Numata M, Matas A J, Simmons R L, Najarian J S. Hepatocellular transplantation in acute liver failure.  Surgery. 1997;  82 124-132
  PubMedGoogle Scholar
• 75 Theise N D, Badve S, Saxena R, Henegariu O, Sell S, Crawford J M, Krause D S. Derivation of hepatocytes from bone marrow cells in mice after radiation-induced myeloablation.  Hepatology. 2000;  31 235-240
  CrossrefPubMedGoogle Scholar
• 76 Watt F M, Hogan B L. Out of Eden: stem cells and their niches.  Science. 2000;  287 1427-1430
  CrossrefPubMedGoogle Scholar
• 77 Wiederkehr J C, Kondos G T, Pollak R. Hepatocyte transplantation for the low-density lipoprotein receptor-deficient state. A study in the Watanabe rabbit.  Transplantation. 1990;  50 466-471
  PubMedGoogle Scholar

Dr. med. Kerstin Leckel

Klinik für Allgemein, Visceral- und Transplantationschirurgie · Charité, Campus Virchow-Klinikum · Universitätsklinikum · Medizinische Fakultät der Humboldt-Universität zu Berlin

Mittelallee 4

13353 Berlin