A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture

Timothy D. Sielaff, Michael Y. Hu, Sridhar Rao, Kristine Groehler, Daidre Olson, Henry J. Mann, Rory P. Remmel, Russell A.D. Shatford, Bruce Amiot, Wei Shou Hu, Frank B. Cerra

Research output: Contribution to journalArticle

56 Citations (Scopus)

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 (3×105cells/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.

Original languageEnglish (US)
Pages (from-to)1459-1463
Number of pages5
JournalTransplantation
Volume59
Issue number10
DOIs
StatePublished - May 1995

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Hepatocytes
Swine
Artificial Liver
Hymecromone
Equipment and Supplies
Egtazic Acid
Serum-Free Culture Media
Collagenases
Collagen Type I
Lidocaine
Cytochrome P-450 Enzyme System
Albumins
Perfusion
Body Weight
Liver

Cite this

Sielaff, T. D., Hu, M. Y., Rao, S., Groehler, K., Olson, D., Mann, H. J., ... Cerra, F. B. (1995). A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture. Transplantation, 59(10), 1459-1463. https://doi.org/10.1097/00007890-199505270-00017

A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture. / Sielaff, Timothy D.; Hu, Michael Y.; Rao, Sridhar; Groehler, Kristine; Olson, Daidre; Mann, Henry J.; Remmel, Rory P.; Shatford, Russell A.D.; Amiot, Bruce; Hu, Wei Shou; Cerra, Frank B.

In: Transplantation, Vol. 59, No. 10, 05.1995, p. 1459-1463.

Research output: Contribution to journalArticle

Sielaff, TD, Hu, MY, Rao, S, Groehler, K, Olson, D, Mann, HJ, Remmel, RP, Shatford, RAD, Amiot, B, Hu, WS & Cerra, FB 1995, 'A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture', Transplantation, vol. 59, no. 10, pp. 1459-1463. https://doi.org/10.1097/00007890-199505270-00017
Sielaff, Timothy D. ; Hu, Michael Y. ; Rao, Sridhar ; Groehler, Kristine ; Olson, Daidre ; Mann, Henry J. ; Remmel, Rory P. ; Shatford, Russell A.D. ; Amiot, Bruce ; Hu, Wei Shou ; Cerra, Frank B. / A technique for porcine hepatocyte harvest and description of differentiated metabolic functions in static culture. In: Transplantation. 1995 ; Vol. 59, No. 10. pp. 1459-1463.
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