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A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture. Transplantation 1995 May 27;59(10):1459-63

Date

05/27/1995

Pubmed ID

7770934

DOI

10.1097/00007890-199505270-00017

Scopus ID

2-s2.0-0029052566   59 Citations

Abstract

Current bioartificial liver devices are based on the use of a large mass of hepatocytes exhibiting differentiated metabolic function. The pig has become a source of interest for the acquisition of such cells-however, harvesting a large mass of highly viable cells has met with difficulty. This study describes a technique for harvesting large quantities of hepatocytes at viabilities greater than 90% and also describes several features documenting differentiated function. Pigs, 6 to 10 kg body weight, underwent in situ two-step whole liver perfusion (ethylene glycol tetraacetic acid and collagenase) and ex vivo cell harvest. Harvests yielded an average of 19.5 billion cells with an average viability of 94.6%. Hepatocytes were then entrapped in type I collagen (3 x 10(5) cells/well) and cultured in serum-free media for 5 days. Pig hepatocytes produced stable amounts of albumin and maintained cytochrome P-450 and glucuronidation activity over 5 days, as shown by the metabolism of lidocaine and 4-methylumbelliferone. These data indicate that pig hepatocytes can be harvested with high yields and can retain viability and differentiated function over at least 5 days of culture, and therefore should prove to be an excellent source of hepatocytes for bioartificial liver devices.

Author List

Sielaff TD, Hu MY, Rao S, Groehler K, Olson D, Mann HJ, Remmel RP, Shatford RA, Amiot B, Hu WS

Author

Sridhar Rao MD, PhD Associate Professor in the Pediatrics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Biotransformation
Cell Separation
Cell Survival
Lidocaine
Liver
Male
Oxygen Consumption
Swine