open access

Vol 45, No 3 (2007)
ORIGINAL PAPERS
Published online: 2007-10-24
Submitted: 2011-12-19
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Effect of anabolics on bovine granulosa-luteal cell primary cultures.

Paola Pregel, Enrico Bollo, Francesca Tiziana Cannizzo, Antonella Rampazzo, Simonetta Appino, Bartolomeo Biolatti
Folia Histochem Cytobiol 2007;45(3):265-271.

open access

Vol 45, No 3 (2007)
ORIGINAL PAPERS
Published online: 2007-10-24
Submitted: 2011-12-19

Abstract

Granulosa cell tumours are observed with increased frequency among calves slaughtered in Northern Italy. The use of illegal anabolics in breeding was taken into account as a cause of this pathology. An in vitro approach was used to detect the possible alterations of cell proliferation induced by anabolics on primary cultures of bovine granulosa-luteal cells. Cultures were treated with different concentrations of substances illegally used in cattle (17beta-estradiol, clenbuterol and boldione). Cytotoxicity was determined by means of MTT test, to exclude toxic effects induced by anabolics and to determine the highest concentration to be tested. Morphological changes were evaluated by means of routine cytology, while PCNA expression was quantified in order to estimate cell proliferation. Cytotoxic effects were revealed at the highest concentrations. The only stimulating effect on cell proliferation was detected in boldione treated cultures: after 48 h treated cells, compared to controls, showed a doubled expression of PCNA. In clenbuterol and 17beta-estradiol treated cells PCNA expression was similar to controls or even decreased. As the data suggest an alteration in cell proliferation, boldione could have a role in the early stage of pathogenesis of granulosa cell tumour in cattle.

Abstract

Granulosa cell tumours are observed with increased frequency among calves slaughtered in Northern Italy. The use of illegal anabolics in breeding was taken into account as a cause of this pathology. An in vitro approach was used to detect the possible alterations of cell proliferation induced by anabolics on primary cultures of bovine granulosa-luteal cells. Cultures were treated with different concentrations of substances illegally used in cattle (17beta-estradiol, clenbuterol and boldione). Cytotoxicity was determined by means of MTT test, to exclude toxic effects induced by anabolics and to determine the highest concentration to be tested. Morphological changes were evaluated by means of routine cytology, while PCNA expression was quantified in order to estimate cell proliferation. Cytotoxic effects were revealed at the highest concentrations. The only stimulating effect on cell proliferation was detected in boldione treated cultures: after 48 h treated cells, compared to controls, showed a doubled expression of PCNA. In clenbuterol and 17beta-estradiol treated cells PCNA expression was similar to controls or even decreased. As the data suggest an alteration in cell proliferation, boldione could have a role in the early stage of pathogenesis of granulosa cell tumour in cattle.
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About this article
Title

Effect of anabolics on bovine granulosa-luteal cell primary cultures.

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 45, No 3 (2007)

Pages

265-271

Published online

2007-10-24

Bibliographic record

Folia Histochem Cytobiol 2007;45(3):265-271.

Authors

Paola Pregel
Enrico Bollo
Francesca Tiziana Cannizzo
Antonella Rampazzo
Simonetta Appino
Bartolomeo Biolatti

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