title: Isolation of rat hepatocytes.
abstract: In vitro models, based on liver cells or tissues, are indispensable in the early preclinical phase of drug development. An important breakthrough in establishing cell models has been the successful high-yield preparation of intact hepatocytes. In this chapter, the practical aspects of the two-step collagenase perfusion method, modified from the original procedure of Seglen, are outlined. Although applicable to the liver of various species, including human, the practical aspects of the method are explained here for rat liver. Critical parameters for the successful isolation of primary rat hepatocytes are highlighted and a troubleshooting guide is provided. In addition, a new development based on the inhibition of histone deacetylase activity is presented. This approach allows inhibition of cell-cycle reentry during hepatocyte isolation, a process known to underlie the dedifferentiation process of cultured hepatocytes.
authors.affiliation: Department of Toxicology, Vrije Universiteit Brussel, Belgium.
authors.forename: Peggy
authors.initials: P
authors.lastname: Papeleu
authors.forename: Tamara
authors.initials: T
authors.lastname: Vanhaecke
authors.forename: Tom
authors.initials: T
authors.lastname: Henkens
authors.forename: Greetje
authors.initials: G
authors.lastname: Elaut
authors.forename: Mathieu
authors.initials: M
authors.lastname: Vinken
authors.forename: Sarah
authors.initials: S
authors.lastname: Snykers
authors.forename: Vera
authors.initials: V
authors.lastname: Rogiers
mesh_terms: D000818:Animals; D002469:Cell Separation; D017364:Collagenases / Q000378:metabolism; D016193:G1 Phase; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D016192:Resting Phase, Cell Cycle
chemical_list: D017364:Collagenases