Proceeding of Veterinary and Animal Science Days 2016, 8th- 10th June, Milan, Italy 

HAF © 2013 

Vol. III, No. 1s  ISSN: 2283-3927 

   

 

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Keywords 

Canine Hepatocyte, BCAA, 

decellularizatio, liver, scaffold 

 

CORRESPONDING AUTHOR 

Matteo Ghiringhelli 

matteo.ghiringhelli@unimi.it 

 

JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

Derivation of canine hepatocyte in vitro 

models to study Branched-Chain Amino Acid 

effects on liver functions. 

 

M. Ghiringhellia*, G. Pennarossaa, A. Di Giancamilloa, S. Brizzolaa, T.A.L.  

Brevinia, C. Domeneghinia, F. Acocellaa, V. Bontempoa 

 

a Department of Health, Animal Science and Food Safety, University of Milan, Via 
Celoria 10, 20133 Milan, Italy 

 

 Abstract 

Branched chain amino acids (BCAA), have been shown to affect human gene expression, protein 

metabolism, apoptosis, and regeneration of hepatocytes. Furthermore, they have been demonstrated 

to inhibit proliferation of liver cancer cells in vitro, and to be essential for lymphocyte proliferation. 

In veterinary medicine, the use of BCAAs as integration of a normal dietary plan, is likely to be a valid 

choice for the same benefit found in human clinical nutrition, although this aspect is still debated. 

Indeed, long-term oral supplementation with BCAAs in the prevention of liver fibrosis and injury in the 

dog's liver is still unclear. Aim of the present study will be to determine how BCAAs preserve liver 

functions in vitro.  To this purpose we have selected and set up three different in vitro models: hepatic 

dog cells and canine hepatocellular carcinoma cells plated in 2D monolayer and hepatic dog cells 

cultured onto 3D scaffolds, obtained from decellularized rabbit liver. All cells adhered and proliferated 

once plated. Cells grown in monolayer quickly entered G0 end arrested growth, ELISA test confirmed 

their ability to produce albumin. Cells grown on scaffold vigorously replicated and showed their 

capability to recellularize ECM rabbit liver. These results, although preliminary, demonstrate that the 

culture conditions used well preserved the original phenotype and function and further support the 

possibility to use in vitro models to successfully study BCAA efficacy in dog.    
 

References 

Boomkens, S.Y., Spee, B., Ljzer, J., Kisjes, R., Egberink, H.F., Van Den Ingh, T.S., Rothuizen, J., Penning, L.C., 2004. The  

establishment and characterization of the first canine hepatocellular carcinoma cell line, which resembles human oncogenic 

expression patterns. Comp Hepatol. 3(1),9. 

Duncan, A.W., Dorrell, C., Grompe, M., 2009. Stem Cells and Liver Regeneration. J Gastroenterol 137(2), 466–81. 

Kruitwagen, H.S., Spee, B., Schotanus, B.A., 2014. Hepatic progenitor cells in canine and feline medicine: potential for 

regenerative strategies. BMC Vet Res. 10,137. 

Rajkumar, R., Victor, R.P., Vinood., B.P., 2015. Branched Chain Amino Acids in Clinical Nutrition, New York, Humana Press.  

Wakshlag, J.J., Kallfelz, F.A., Wakshlag, R.R., Davenport, G.M., 2006. The effects of branched-chain amino acids on canine 

neoplastic cell proliferation and death. J Nutr. 136(7), 2007S-2010S. 

http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en