Culture of bovine hepatocytes: a non-perfusion technique for cell isolation

Spotorno VG1, Hidalgo A, Barbich M, Lorenti A, Zabal O.
Source: Cytotechnology
Publication Date: (2006)
Issue: 51(2): 51-56
Cells used in publication:
Hepatocyte, mouse
Species: mouse
Tissue Origin: liver
Hepatocyte, rat
Species: rat
Tissue Origin: liver
Hepatocyte, human
Species: human
Tissue Origin: liver
Hepatocyte, Cynomolgus
Species: monkey
Tissue Origin: liver
Hepatocyte, Rhesus
Species: monkey
Tissue Origin: liver
Hepatocyte, canine
Species: canine
Tissue Origin:
In this work we have studied the isolation and culture of mature bovine hepatocytes on plastic dishes without exogenous matrix. The liver has been disaggregated in a collagenase solution instead of undergoing a perfusion step. After a few days in culture, the plates showed several clusters of different cell types. Although the average yield was 1.60+/-0.57x10(8) viable liver cells per gram of tissue, these cultures were formed by non-parenchymal cells and only very few or none by parenchymal cells. In these cultures, actin structures used as a marker for Stellate (Ito) cells have been visualized by immunocytochemical techniques. In order to increase the proportion of parenchymal cells a centrifugation on Percoll, which separates cell sub-populations, has been introduced. Though the yield was lower than in the previous method, these pre-purified cultures were only composed of hepatocytes. It has been shown that these cells exhibited albumin synthesis, which is a specific hepatocytes function. In addition, these cultures were capable of producing metabolites of 7-ethoxycoumarin at a higher rate than non purified cell cultures. Therefore this simplified procedure for the isolation and culture of functional and viable hepatocytes may be applied for in vitro studies in bovine.