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Cytotechnology. 2006 Jun;51(2):51-6. doi: 10.1007/s10616-006-9000-0. Epub 2006 Sep 5.

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

Author information

1
Instituto Tecnología de Alimentos. C.I.A., Instituto Nacional de Tecnología Agropecuaria (INTA), c.c. 77 B1708WAB. Morón, Buenos Aires, Argentina, vspotorno@cnia.inta.gov.ar.

Abstract

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.

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