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. 1986 Sep;204(3):259–271. doi: 10.1097/00000658-198609000-00005

New method of hepatocyte transplantation and extracorporeal liver support.

A A Demetriou, J Whiting, S M Levenson, N R Chowdhury, R Schechner, S Michalski, D Feldman, J R Chowdhury
PMCID: PMC1251274  PMID: 3530153

Abstract

A technique has been developed by the authors that allows hepatocyte attachment on collagen-coated microcarriers resulting in prolonged hepatocyte viability and function both in vivo and in vitro. Rat hepatocytes were obtained by portal vein collagenase perfusion. Intraperitoneally transplanted microcarrier-attached normal hepatocytes into congeneic Gunn rats were functioning 3-4 weeks later, as shown by the presence and persistence of conjugated bilirubin in recipient bile, sustained decrease in serum bilirubin, uptake of Tc99m-DESIDA, and morphologic criteria. Intraperitoneal transplantation of normal microcarrier-attached hepatocytes into genetically albumin deficient rats (NAR) resulted in marked increase in plasma albumin levels (6 days without and 21 days with Cyclosporin A immunosuppression). Microcarrier-attached hepatocytes transplanted after 2 weeks of storage at -80 C into congeneic Gunn rats were viable and functional as assessed by criteria outlined above. An extracorporeal liver perfusion system was developed using the microcarrier-attached hepatocytes that was capable of synthesizing and conjugating bilirubin and synthesizing liver-specific proteins.

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264FIGS. 5A and B.
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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abouna G. M., Fisher L. M., Still W. J., Hume D. M. Acute hepatic coma successfully treated by extracorporeal baboon liver perfusions. Br Med J. 1972 Jan 1;1(5791):23–25. doi: 10.1136/bmj.1.5791.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrett P. V., Mullins F. X., Berlin N. I. Studies on the biosynthesis production of bilirubin-C14: an improved method utilizing delta-aminolevulinic acid-4-C14 in dogs. J Lab Clin Med. 1966 Dec;68(6):905–912. [PubMed] [Google Scholar]
  3. Baumgartner D., LaPlante-O'Neill P. M., Sutherland D. E., Najarian J. S. Effects of intrasplenic injection of hepatocytes, hepatocyte fragments and hepatocyte culture supernatants on D-galactosamine-induced liver failure in rats. Eur Surg Res. 1983;15(3):129–135. doi: 10.1159/000128344. [DOI] [PubMed] [Google Scholar]
  4. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blanckaert N., Kabra P. M., Farina F. A., Stafford B. E., Marton L. J., Schmid R. Measurement of bilirubin and its monoconjugates and diconjugates in human serum by alkaline methanolysis and high-performance liquid chromatography. J Lab Clin Med. 1980 Aug;96(2):198–212. [PubMed] [Google Scholar]
  6. Brunner G., Holloway C. J., Lösgen H. Large agarose beads for extracorporeal detoxification systems. Preparation and enzymatic properties of agarose-bound UDP-glucuronyltransferase. Int J Artif Organs. 1979 May;2(3):163–169. [PubMed] [Google Scholar]
  7. Chowdhury J. R., Chowdhury N. R., Wu G., Shouval R., Arias I. M. Bilirubin mono- and diglucuronide formation by human liver in vitro: assay by high-pressure liquid chromatography. Hepatology. 1981 Nov-Dec;1(6):622–627. doi: 10.1002/hep.1840010610. [DOI] [PubMed] [Google Scholar]
  8. Contini S., Pezzarossa A., Sansoni P., Mazzoni M. P., Botta G. C. Hepatocellular transplantation in rats with toxic induced liver failure: results of iso-, allo- and xenografts. Ital J Surg Sci. 1983;13(1):25–30. [PubMed] [Google Scholar]
  9. Eiseman B., Liem D. S., Raffucci F. Heterologous liver perfusion in treatment of hepatic failure. Ann Surg. 1965 Sep;162(3):329–345. doi: 10.1097/00000658-196509000-00003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eiseman B., Norton L., Kralios N. C. Hepatocyte perfusion within a centrifuge. Surg Gynecol Obstet. 1976 Jan;142(1):21–28. [PubMed] [Google Scholar]
  11. Gebb C., Clark J. M., Hirtenstein M. D., Lindgren G., Lindskog U., Lundgren B., Vretblad P. Alternative surfaces for microcarrier culture of animal cells. Dev Biol Stand. 1981;50:93–102. [PubMed] [Google Scholar]
  12. Gjessing R., Seglen P. O. Adsorption, simple binding and complex binding of rat hepatocytes to various in vitro substrata. Exp Cell Res. 1980 Sep;129(1):239–249. doi: 10.1016/0014-4827(80)90347-x. [DOI] [PubMed] [Google Scholar]
  13. Groth C. G., Arborgh B., Björkén C., Sundberg B., Lundgren G. Correction of hyperbilirubinemia in the glucuronyltransferase-deficient rat by intraportal hepatocyte transplantation. Transplant Proc. 1977 Mar;9(1):313–316. [PubMed] [Google Scholar]
  14. Harder F., Floersheim G. L., Jeekel J., van den Linden C., Mihatsch J., Rüedi T., Tondelli P. Prolongation of canine renal allografts: plasma or liver instead of blood transfusions combined with preoperative nonspecific immunosuppression. Transplant Proc. 1979 Mar;11(1):978–981. [PubMed] [Google Scholar]
  15. Hay F. C., Nineham L. J., Roitt I. M. Routine assay for the detection of immune complexes of known immunoglobulin class using solid phase C1q. Clin Exp Immunol. 1976 Jun;24(3):396–400. [PMC free article] [PubMed] [Google Scholar]
  16. Heirwegh K. P., Van Hees G. P., Leroy P., Van Roy F. P., Jansen F. H. Heterogeneity of bile pigment conjugates as revealed by chromatography of their ethyl anthranilate azopigments. Biochem J. 1970 Dec;120(4):877–890. doi: 10.1042/bj1200877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Howe R. B., Berk P. D., Bloomer J. R., Berlin N. I. Preparation and properties of specifically labeled radiochemically stable 3H-bilirubin. J Lab Clin Med. 1970 Mar;75(3):499–502. [PubMed] [Google Scholar]
  18. Hughes R. D., Ton H. Y., Langley P. G., Silk D. B., Williams R. The use of an in vitro haemoperfusion circuit to evaluate the blood compatibility of albumin-coated Amberlite XAD-7 resin. Int J Artif Organs. 1978 May;1(3):129–134. [PubMed] [Google Scholar]
  19. Kusano M., Mito M. Observations on the fine structure of long-survived isolated hepatocytes inoculated into rat spleen. Gastroenterology. 1982 Apr;82(4):616–628. [PubMed] [Google Scholar]
  20. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  21. Leffert H. L., Koch K. S., Moran T., Rubalcava B. Hormonal control of rat liver regeneration. Gastroenterology. 1979 Jun;76(6):1470–1482. [PubMed] [Google Scholar]
  22. Makowka L., Falk R. E., Rotstein L. E., Falk J. A., Nossal N., Langer B., Blendis L. M., Phillips M. J. Cellular transplantation in the treatment of experimental hepatic failure. Science. 1980 Nov 21;210(4472):901–903. doi: 10.1126/science.7001630. [DOI] [PubMed] [Google Scholar]
  23. Makowka L., Falk R. E., Rotstein L. E., Falk J. A., Nossal N., Langer B., Blendis L. M., Phillips M. J. Reversal of experimental acute hepatic failure in the rat. J Surg Res. 1980 Dec;29(6):479–487. doi: 10.1016/0022-4804(80)90016-5. [DOI] [PubMed] [Google Scholar]
  24. Makowka L., Rotstein L. E., Falk R. E., Falk J. A., Langer B., Nossal N. A., Blendis L. M., Phillips M. J. Reversal of toxic and anoxic induced hepatic failure by syngeneic, allogeneic, and xenogeneic hepatocyte transplantation. Surgery. 1980 Aug;88(2):244–253. [PubMed] [Google Scholar]
  25. Makowka L., Rotstein L. E., Falk R. E., Falk J. A., Zuk R., Langer B., Blendis L. M., Phillips M. J. Studies into the mechanism of reversal of experimental acute hepatic failure by hepatocyte transplantation. 1. Can J Surg. 1981 Jan;24(1):39–44. [PubMed] [Google Scholar]
  26. Matas A. J., Sutherland D. E., Steffes M. W., Mauer S. M., Sowe A., Simmons R. L., Najarian J. S. Hepatocellular transplantation for metabolic deficiencies: decrease of plasms bilirubin in Gunn rats. Science. 1976 May 28;192(4242):892–894. doi: 10.1126/science.818706. [DOI] [PubMed] [Google Scholar]
  27. Ohkawa M., Hayashi H., Chaudry I. H., Clemens M. G., Baue A. E. Effects of regenerating liver cytosol on drug-induced hepatic failure. Surgery. 1985 Apr;97(4):455–462. [PubMed] [Google Scholar]
  28. Patterson M. K., Jr Measurement of growth and viability of cells in culture. Methods Enzymol. 1979;58:141–152. doi: 10.1016/s0076-6879(79)58132-4. [DOI] [PubMed] [Google Scholar]
  29. Reid L. M., Rojkind M. New techniques for culturing differentiated cells: reconstituted basement membrane rafts. Methods Enzymol. 1979;58:263–278. doi: 10.1016/s0076-6879(79)58142-7. [DOI] [PubMed] [Google Scholar]
  30. Seglen P. O. Preparation of isolated rat liver cells. Methods Cell Biol. 1976;13:29–83. doi: 10.1016/s0091-679x(08)61797-5. [DOI] [PubMed] [Google Scholar]
  31. Sommer B. G., Sutherland D. E., Matas A. J., Simmons R. L., Najarian J. S. Hepatocellular transplantation for treatment of D-galactosamine-induced acute liver failure in rats. Transplant Proc. 1979 Mar;11(1):578–584. [PubMed] [Google Scholar]
  32. Sommer B. G., Sutherland D. E., Simmons R. L., Najarian J. S. Hepatocellular transplantation for experimental acute liver failure in dogs. Surg Forum. 1979;30:279–281. [PubMed] [Google Scholar]
  33. Sommer B. G., Sutherland D. E., Simmons R. L., Najarian J. S. Hepatocellular transplantation for experimental ischemic acute liver failure in dogs. J Surg Res. 1980 Oct;29(4):319–325. doi: 10.1016/0022-4804(80)90064-5. [DOI] [PubMed] [Google Scholar]
  34. Starzl T. E., Klintmalm G. B., Iwatsuki S., Fernandez-Bueno C. Past and future prospects of orthoptic liver transplantation. Arch Surg. 1981 Oct;116(10):1342–1343. doi: 10.1001/archsurg.1981.01380220086013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Toledo-Pereyra L. H., Gordon D. A., MacKenzie G. H. Immunologic response to liver cell allografts. Am Surg. 1982 Jan;48(1):28–31. [PubMed] [Google Scholar]
  36. Trotman B. W., Roy-Chowdhury J., Wirt G. D., Bernstein S. E. Azodipyrroles of unconjugated and conjugated bilirubin using diazotized ethyl anthranilate in dimethyl sulfoxide. Anal Biochem. 1982 Mar 15;121(1):175–180. doi: 10.1016/0003-2697(82)90572-3. [DOI] [PubMed] [Google Scholar]
  37. Vroemen J. P., Blanckaert N., Buurman W. A., Heirwegh K. P., Kootstra G. Treatment of enzyme deficiency by hepatocyte transplantation in rats. J Surg Res. 1985 Sep;39(3):267–275. doi: 10.1016/0022-4804(85)90152-0. [DOI] [PubMed] [Google Scholar]
  38. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  39. Wolf C. F., Minick C. R., McCoy C. H. Morphologic examination of a prototype liver assist device composed of cultured cells and artificial capillaries. Int J Artif Organs. 1978 Jan;1(1):45–51. [PubMed] [Google Scholar]
  40. Woods R. J., Fuller B. J., Attenburrow V. D., Nutt L. H., Hobbs K. E. Functional assessment of hepatocytes after transplantation into rat spleen.. Transplantation. 1982 Feb;33(2):123–126. doi: 10.1097/00007890-198202000-00004. [DOI] [PubMed] [Google Scholar]
  41. Woods R. J., Parbhoo S. P. An explanation for the reduction in bilirubin levels in congenitally jaundiced Gunn rats after transplantation of isolated hepatocytes. Eur Surg Res. 1981;13(4):278–284. doi: 10.1159/000128194. [DOI] [PubMed] [Google Scholar]
  42. ten Berg R. G., Ernst P., van Maldegem-Dronkers C., Marquet R., Westbroek D. L. Effect of viable isolated hepatocytes or hepatocyte fractions on survival rate following galactosamine-induced acute liver failure. Eur Surg Res. 1985;17(2):109–118. doi: 10.1159/000128455. [DOI] [PubMed] [Google Scholar]

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