Details

Autor(en) / Beteiligte

• Kwan, I. (McGill University, Ste Anne de Bellevue, Canada.)
• Farookhi, R
• Hung The Huynh
• Murphy, B.D
• Turner, J.D
• Downey, B.R

Titel

Steroidogenic properties of a spontaneously established porcine granulosa cell line (PGC-2)

Ist Teil von

• Molecular reproduction and development, 1996-11, Vol.45 (3), p.299-307

Ort / Verlag

Hoboken: Wiley Subscription Services, Inc., A Wiley Company

Erscheinungsjahr

1996

Quelle

Wiley-Blackwell Journals

Beschreibungen/Notizen

• A spontaneously established porcine granulosa cell line (PGC‐2) was cloned through the continuous culturing of primary granulosa cells collected from equine chorionic gonadotropin (eCG)‐treated prepubertal gilts. This established cell line has undergone ∼︁ 100 passages and shows contact‐inhibition of growth. PGC‐2 stained with a monoclonal antibody (mAb) directed against cytokeratin, indicating its epithelial nature, but not with a mAb directed against vimentin, suggesting that it is not fibroblast derived. Immunoblotting revealed that PGC‐2 expresses cadherin, an epithelial Ca??? dependent cell adhesion molecule. The cells were dependent on serum for growth and had a doubling time of ∼︁ 20 hr when cultured with 10% fetal bovine serum. The cell line was examined for the presence of FSH receptors, cAMP responses, and steroidogenic capabilities. The cell line lacks FSH receptors as assessed by radiolabelled‐ligand binding, and no transcripts for FSH receptor were detected by Northern blotting of total cellular RNA. Neither FSH nor cholera toxin (0.5 ng/mL) stimulated increases in cAMP levels in these cells, whereas forskolin 10 μM) induced a fivefold increase in cAMP production. When a higher concentration of cholera toxin (300 ng/mL) was used, however, cAMP levels doubled by 2 hr. Despite a lack of responsiveness to purified oFSH or oLH, the cells were capable of progesterone and estradiol production when provided with the appropriate substrates. We conclude that PGC‐2 display properties that are similar to immature granulosa cells and may provide a suitable in vitro model for the study of granulosa cell function. © 1996 Wiley‐Liss, Inc.