Isolation and cultivation of adult primary bovine hepatocytes from abattoir derived liver

Brief Research Report: Effect of Triiodothyronine on Hepatic Growth Hormone Receptor Expression in Primary Bovine Hepatocytes

“…We could confirm that there is an additive effect of T3 on the expression of GHR and IGF 1 mRNA in primary bovine hepatocytes, using a well-established cell culture model (12)(13)(14) with primary hepatocytes derived from an adult dairy cow. A pivotal question is how T3 can induce IGF-1 production in primary hepatocytes without growth hormone (GH) being added to the cell culture medium.…”

“…Expression of GHR and IGF 1 mRNA was determined using a quantitative reverse transcriptase PCR. Total RNA was extracted using TRIzolTM Reagent (REF 15596018; Life Technologies Corporation, Carlsbad, USA), as previously published by Ehrhardt and Schmicke (13). The concentration and purity of extracted RNA were measured using a NanoDrop (NanoDrop 2000c, Thermo Fisher Scientific, Waltham, Massachusetts, USA) before transcription of RNA into complementary DNA (cDNA).…”

“…PCR was performed using the Rotor-Gene Q (QUIAGEN GmbH, Hilden, Germany). Primers for GHR1A and IGF-1 were previously used and validated (12,13).…”

Tumor necrosis factor alpha and palmitate simulate bovine fatty liver disease in vitro when using abattoir-derived primary bovine hepatocytes isolated by a novel nonperfusion method

“…A few in vitro culture methods for bovine hepatic cells have been presented, but they are complicated in several ways because they involve one or more of the following: collagenase perfusion ( Berry and Friend, 1969 ; Witte et al, 2019 ) or complex culture media ( Ehrhardt and Schmicke, 2016 ) often supplemented with exogenous growth factors or steroids ( Giantin et al, 2012 ). A second consideration is that most previous research used male calves ( Donkin and Armentano, 1993 ) because of availability and cost, or female newborn calves ( Li et al, 2018 ) that are more costly and may not represent the physiology of the adult cow, and additionally will have to be euthanized.…”

“…A second consideration is that most previous research used male calves ( Donkin and Armentano, 1993 ) because of availability and cost, or female newborn calves ( Li et al, 2018 ) that are more costly and may not represent the physiology of the adult cow, and additionally will have to be euthanized. Moreover, abattoir-derived liver has been utilized for primary cell culture in several species such as equine ( Stefanski et al, 2013 ), porcine ( Gerlach et al, 1993 ), and bovine species ( Panda et al, 2015 ; Ehrhardt and Schmicke, 2016 ). Because of the lack of available established cell lines or established simple primary cell culture methodologies and with the need for further mechanistic studies in mind, we sought to develop an in vitro model of bovine hepatic lipidosis.…”