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An immortalized gonadotropic cell line could not be developed. Functional lifetime of culture was extended by IGF-1

Already available in vitro methods developed to monitor the estrogenic activity of EEs only provide data reflecting estrogen receptor binding or mitogenic activity. Because of the pivotal role of GTHs within the BPG axis, it is essential to determine the effect of EEs on GnRH receptors and GTH synthesis and release. This can be achieved by experiments as outlined in the present research program. Although such an approach is necessary at the beginning, it is time-consuming and specialized expertise and suitable aquarium facilities are needed. Therefore, in the present program, it was attempted to develop an immortalized gonadotroph cell line, to be made available for future research on the cellular and molecular mechanisms of action of EEs (or other pollutants) in gonadotrophs. We did not succeed in the development of an immortalized gonadotroph cell line. The main obstacle was the terminally differentiated state of the gonadotroph cells even in juvenile fish. Trials to stimulate cells to re-enter cell cycling failed. Hence, the immortalisation strategy, introduction of an oncogene, remains ineffective.

We succeeded instead by adding IGF-1 to the culture medium, to extend the functional survival period of gonadotrophs in primary culture to 3 weeks.

This result can be used in similar research projects on the effects of potential endocrine disrupting agents on pituitary gonadotrophs.

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Reported by

University of Utrecht
Padualaan 8
3584 CH Utrecht
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