Skip to main content
Journal cover image

Isolation and characterization of biliary epithelial cells from rainbow trout liver.

Publication ,  Journal Article
Blair, JB; Ostrander, GK; Miller, MR; Hinton, DE
Published in: In vitro cellular & developmental biology. Animal
November 1995

Lectin binding and density gradient centrifugation were explored for isolating epithelial cells from trout liver. Hepatocytes exhibited preferential attachment of coverslips coated with Phaseolus vulgaris erythroagglutinin. Biliary epithelial cells attached with glycine max agglutinin; however, significant attachment of cellular debris limited the use of glycine max agglutinin. Percoll-density gradient centrifugation separated liver cells into two distinct populations with biliary cells and hepatocytes banding at densities of 1.04 and 1.09, respectively. A discontinuous gradient composed of 13% Ficoll (wt/wt) separated biliary cells from hepatocytes. The recovery of highly enriched biliary epithelial cells from trout liver using Ficoll gradients yielded approximately 8 million cells (0.1 ml packed cells) from 10 g liver. Western blot analysis demonstrated that the cytokeratin profile for extracts from biliary epithelial cell-enriched populations differ significantly from those seen with whole liver extracts or with extracts with hepatocyte-enriched populations. Ficoll-gradient purified biliary cells and hepatocytes attached to culture plates coated with trout skin extract and carried out linear incorporation of leucine into protein and thymidine into DNA for 24 h. A mixture of growth hormones (insulin, epidermal growth factor, and dexamethasone) stimulated thymidine incorporation into DNA; however, long-term culture of dividing biliary epithelial cells was not achieved. Chemical analysis of neutral and acidic glycolipids indicated that hepatocytes and biliary cells have similar glycolipid profiles with an exception in the region of GM3 mobility, which is attributed to differences in the ceramide moiety. These studies provide a starting point for further characterization of unique cell types of the trout liver that may be important in their responses to toxic and carcinogenic agents.

Duke Scholars

Published In

In vitro cellular & developmental biology. Animal

DOI

EISSN

1543-706X

ISSN

1071-2690

Publication Date

November 1995

Volume

31

Issue

10

Start / End Page

780 / 789

Related Subject Headings

  • beta-Galactoside alpha-2,3-Sialyltransferase
  • Sialyltransferases
  • Protein Binding
  • Oncorhynchus mykiss
  • Liver
  • Lectins
  • Keratins
  • Glycolipids
  • Centrifugation
  • Cells, Cultured
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Blair, J. B., Ostrander, G. K., Miller, M. R., & Hinton, D. E. (1995). Isolation and characterization of biliary epithelial cells from rainbow trout liver. In Vitro Cellular & Developmental Biology. Animal, 31(10), 780–789. https://doi.org/10.1007/bf02634120
Blair, J. B., G. K. Ostrander, M. R. Miller, and D. E. Hinton. “Isolation and characterization of biliary epithelial cells from rainbow trout liver.In Vitro Cellular & Developmental Biology. Animal 31, no. 10 (November 1995): 780–89. https://doi.org/10.1007/bf02634120.
Blair JB, Ostrander GK, Miller MR, Hinton DE. Isolation and characterization of biliary epithelial cells from rainbow trout liver. In vitro cellular & developmental biology Animal. 1995 Nov;31(10):780–9.
Blair, J. B., et al. “Isolation and characterization of biliary epithelial cells from rainbow trout liver.In Vitro Cellular & Developmental Biology. Animal, vol. 31, no. 10, Nov. 1995, pp. 780–89. Epmc, doi:10.1007/bf02634120.
Blair JB, Ostrander GK, Miller MR, Hinton DE. Isolation and characterization of biliary epithelial cells from rainbow trout liver. In vitro cellular & developmental biology Animal. 1995 Nov;31(10):780–789.
Journal cover image

Published In

In vitro cellular & developmental biology. Animal

DOI

EISSN

1543-706X

ISSN

1071-2690

Publication Date

November 1995

Volume

31

Issue

10

Start / End Page

780 / 789

Related Subject Headings

  • beta-Galactoside alpha-2,3-Sialyltransferase
  • Sialyltransferases
  • Protein Binding
  • Oncorhynchus mykiss
  • Liver
  • Lectins
  • Keratins
  • Glycolipids
  • Centrifugation
  • Cells, Cultured