Survival, proliferation, and functions of porcine hepatocytes encapsulated in coated alginate beads: a step toward a reliable bioartificial liver.
Orthotopic liver transplantation is the most effective treatment for fulminant hepatic failure. As an alternative treatment, an efficient extracorporeal bioartificial liver should contain a large yield of functional hepatocytes with an immunoprotective barrier, for providing temporary adequate metabolic support to allow spontaneous liver regeneration or for acting as a bridge toward transplantation. Survival, proliferation, and functions of porcine hepatocytes were evaluated in primary cultures and after embedding in alginate beads, which were subsequently coated with a membrane made by a transacylation reaction between propylene glycol alginate and human serum albumin. Disruption of total pig livers by collagenase perfusion/recirculation allowed the obtention of up to 10(11) hepatocytes with a viability greater than 95%. Hepatocytes in conventional cultures or embedded in coated alginate beads survived for about 10 days, secreted proteins, particularly albumin, and maintained several phase I and II enzymatic activities, namely ethoxyresorufin-O-deethylase, oxidation of nifedipine to pyridine, phenacetin deethylation to paracetamol, glucuroconjugation of paracetamol, and N-acetylation of procainamide. Typical features of mitosis and [3H]thymidine incorporation indicated that porcine hepatocytes proliferated in both conventional cultures and alginate beads. The efficacy of the membrane surrounding alginate beads for protecting cells from immunoglobulins was tested by embedding HLA-typed human lymphocytes, which were subsequently incubated with specific anti-HLA immunoglobulin G and complement. These data show that large yields of porcine hepatocytes that are embedded in coated alginate beads remain functional and are isolated from large molecular weight molecules, such as immunoglobulins. This system represents a promising tool for the design of an extracorporeal bioartificial liver, containing xenogeneic hepatocytes, to treat acute liver disease in humans.
Joly
A
A
Detoxication and Tissue Repair Unit 456, Institut National de la Santéde la Recherche Médicale, Rennes I University School of Medicine, France.
Desjardins
J F
JF
Fremond
B
B
Desille
M
M
Campion
J P
JP
Malledant
Y
Y
Lebreton
Y
Y
Semana
G
G
Edwards-Levy
F
F
Levy
M C
MC
Clement
B
B
D000082:Acetaminophen / Q000493:pharmacokinetics; D000464:Alginates; D000818:Animals; D001711:Biotransformation; D002214:Capsules; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D005260:Female; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D008107:Liver Diseases / Q000628:therapy; D019164:Liver, Artificial; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D011342:Procainamide / Q000493:pharmacokinetics; D014176:Protein Biosynthesis; D012709:Serum Albumin / Q000096:biosynthesis; D013552:Swine
D000464:Alginates; D002214:Capsules; D006603:Hexuronic Acids; D012709:Serum Albumin; D000082:Acetaminophen; D020723:Glucuronic Acid; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D011342:Procainamide