title: Acellular liver matrix improves the survival and functions of isolated rat hepatocytes cultured in vitro.
abstract: Alternative approaches to overcome the shortage of donors for liver transplantation may be the use of hepatocytes for bioartificial devices or transplantation. Therefore, the setting-up of new in vitro culture techniques allowing the long-term survival and functional maintenance of hepatocytes represents a formidable challenge. Aim of this study was to obtain a liver homologous acellular matrix (HAM) able to support viability and metabolic functions of rat hepatocytes in primary culture. HAMs were prepared by sequential incubation of rat liver slices in deoxycholic acid and DNase solutions. Dispersed rat hepatocytes were obtained by collagenase digestion and mechanical disaggregation. Isolated hepatocytes were seeded on uncoated and collagen- or HAM-coated tissue culture plastic wells. Cultures were examined by scanning electron microscopy (SEM), and the viability of hepatocytes and their ability to produce albumin and urea were assessed. The viability of freshly dispersed hepatocytes was about 98%. Hepatocytes seeded on HAM exhibited a significantly higher viability and a markedly lower apoptotic rate than those grown on plastic or collagen. Accordingly, albumin and urea nitrogen productions were significantly higher in HAM-cultured hepatocytes. SEM showed that hepatocytes seeded on HAM displayed a clustered organization, and were well anchored to the matrix and morphologically stable. Taken together, these findings indicate that HAM strongly improves viability and functional activity of rat hepatocytes cultured in vitro.
authors.affiliation: Department of Surgical and Gastroenterological Sciences, Section of Gastroenterology, University of Padua, Via Gabelli 65, I-35121 Padua, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Burra
authors.forename: Silvia
authors.initials: S
authors.lastname: Tomat
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Conconi
authors.forename: Carlo
authors.initials: C
authors.lastname: Macchi
authors.forename: Francesco P
authors.initials: FP
authors.lastname: Russo
authors.forename: Pier Paolo
authors.initials: PP
authors.lastname: Parnigotto
authors.forename: Remo
authors.initials: R
authors.lastname: Naccarato
authors.forename: Gastone G
authors.initials: GG
authors.lastname: Nussdorfer
mesh_terms: D000418:Albumins / Q000096:biosynthesis / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D046508:Culture Techniques; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008099:Liver / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D008110:Liver Extracts / Q000494:pharmacology*; D008297:Male; D008855:Microscopy, Electron, Scanning; D051381:Rats; D017207:Rats, Sprague-Dawley; D014508:Urea / Q000378:metabolism
chemical_list: D000418:Albumins; D008110:Liver Extracts; D014508:Urea