title: A simple method for the simultaneous isolation of stellate cells and hepatocytes from rat liver tissue.
abstract: Hepatic stellate cells (HSCs), also referred to as Ito cells, perisinusiodal cells and fat-storing cells, have numerous vital functions. They are the main extracellular matrix-producing cells within the liver and are involved in the storage of retinol. HSCs are also known to secrete a number of liver mitogens. Current isolation techniques are cumbersome and most require a pronase digestion step, which destroys any hepatocytes present. We present a simple method for isolation and culture of hepatic stellate cells from the normally discarded washings from a two-step collagenase hepatocyte isolation, which has shown a yield of more than 1.5 x 10(6) viable HSCs after 5 days in culture. The cells were positively identified as HSCs by staining for two intermediate filaments (desmin and GFAP) and observing their distinct morphology from other liver cell types. This efficient method allows rapid and consistent isolation of stellate cells to give a culture that may be passaged several times.
authors.affiliation: School of Pharmaceutical Sciences, University of Nottingham, Nottingham, UK.
authors.forename: Lisa
authors.initials: L
authors.lastname: Riccalton-Banks
authors.forename: Rena
authors.initials: R
authors.lastname: Bhandari
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Fry
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Shakesheff
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D017364:Collagenases / Q000378:metabolism; D003893:Desmin / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology*; D050356:Lipid Metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D008858:Microscopy, Phase-Contrast; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014801:Vitamin A / Q000378:metabolism
chemical_list: D003893:Desmin; D005904:Glial Fibrillary Acidic Protein; D014801:Vitamin A; D017364:Collagenases