Endocrine Abstracts (2004) 7 P196

Biochemical evidence for follicular maturation (2): production of progesterone, inhibins and activin-A by granulosa cells from size matched follicles

XS Wen, AJ Tozer, SA Butler & RK Iles

1Williamson Laboratory, St Barts Hospital, Queen Mary Medical School, London, UK.

Background: Being little known about the biochemistry behind the follicular content or granulosa cells surrounding a mature oocyte in Assisted Reproductive Technology, our study set up to investigate the changes in steroid and cytokines' productions by cultured granulosa cells (GC) taken from individual follicles from women undergoing Controlled Ovarian Hyperstimulation (COH).

MATERIALS AND METHODS: Granulosa cells isolated from 75 individual follicles were cultured up to 72 hours. Progesterone, inhibin-A, inhibin-B and activin-A contents after 24 hours and 72 hours continuous culture were detected by commercialized Enzyme-linked Immunosorbent Assay (ELISA).

RESULTS: Inhibin A, inhibin B and progesterone continued to be secreted by GCs in vitro, Activin A levels were only marginally detected in 50% of cultures. Progesterone production was highest from GCs isolated from large follicles and increased proportionally in all cultures when stimulated by hCG. Basal inhibin A production was highest from cells isolated from large follicles. However hCG had no effect on Inhibin A secretion. Basal inhibin-B productions by GC had no correlation with the size of follicle from which they were originated. However, hCG stimulation not only decreased inhibin B production, but this effect was most marked in GC isolated from larger follicles.

CONCLUSION: Post ovulation granulosa cells differentiate into luteal cells. Our in vitro cultures of granulosa cells from follicles somewhat mimicks this biological phenomenon. Production of Inhibins and Activins were surprisingly different from the patterns seen in follicular fluid. Activin A was not to be actively produced. In addition one marked effect of luteal differentiation would appear to be the marked decrease in inhibin B production in response to hCG stimulation.

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