Fol. Biol. 2015, 61, 184-194

https://doi.org/10.14712/fb2015061050184

Characterization of Porcine Granulosa Cell Line AVG-16

A. Sadowska1, A. Nynca2, M. Korzeniewska1, J. Piasecka-Srader3, M. Jablonska1, K. Orlowska1, S. Swigonska2, Renata E. Ciereszko1,2

1Department of Animal Physiology, University of Warmia and Mazury, Olsztyn, Poland
2Laboratory of Molecular Diagnostics, University of Warmia and Mazury, Olsztyn, Poland
3Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS, USA

Received May 2015
Accepted June 2015

Commercially available, but not yet characterized, the AVG-16 granulosa cell line was established from granulosa cells of medium porcine follicles. To examine the suitability of the AVG-16 cell line for studying the molecular mechanism of action of various environmental oestrogens, we investigated: 1/ cell morphology (by standard haematoxylin and eosin (HE) staining); 2/ basal and follicle-stimulating hormone (FSH) or luteinizing hormone (LH) -stimulated steroid hormone (progesterone; P4 and 17β-oestradiol; E2) secretion (by radioimmunoassay) and 3/ expression of receptors involved in the regulation of granulosa cell function: FSH receptor (FSHR), LH receptor (LHR), oestrogen receptor α (ERα), oestrogen receptor β (ERβ) and aryl hydrocarbon receptor (AhR). mRNA and protein expression was determined by RT-PCR and fluorescence immunocytochemistry, respectively. The secretion of P4 and E2 by AVG-16 cells was in the range of steroid hormone secretion by porcine cultured primary granulosa cells. Neither FSH (100 ng/ml) nor LH (100 ng/ml) affected P4 and E2 secretion by AVG-16 cells. The presence of FSHR and LHR at both mRNA and protein level was not demonstrated in the cells. However, AVG-16 cells were found to express mRNA and protein of ERα, ERβ and AhR. The results of our study showed that AVG-16 cells possess the capability of steroid hormone production, and both oestrogen receptors and AhR are present in these cells. Therefore, AVG-16 cells may serve as an unlimited source of homogenous porcine granulosa cells useful for studying the effects of environmental oestrogens on ovarian physiology.

Funding

This study was supported by The Ministry of Science and Higher Education in Poland (2012/05/B/NZ9/03333, UWM No. 528.0206.0806.).

References

65 live references