Reproduction Abstracts

Background: As a consequence of cryopreservation of human ovaries preformed in our laboratory we have access to human small antral follicles.

Material: Human granulosa cells (GC) from individual small antral follicles (4–6 mm in diameter) were spilt in two and either cultured or snap-frozen immediately after isolation from the follicular fluid. Microarray analysis was used on both the cultured and the non-cultured sample, to examine their gene expression profiles. Ten individual follicles, from six women were used in the present study. The results of the microarray analysis were validated on qPCR.

Results: Combining culture studies, microarray-and qPCR-analysis major changes in the gene expression profiles were observed between cultured and non-cultured GC samples.

The non-cultured samples showed a gene expression profile with high expression of FSH receptor (FSHR), anti-Müllerian hormone (AMH) and androgen receptor (AR), and low expression levels of the CYP19a1 and LH receptor (LHR), corresponding with what is expected in GC from human small antral follicles. Controversially, the GC from the cultured samples showed a gene expression profile with significantly lower FSHR, AMH, and AR expression and significantly higher expression of CYP19a1 and LHR, corresponding with expression profiles of GC from preovulatory follicles. Further, several other genes characteristic for GC from preovulatory follicles were up-regulated in the cultured cells.

Conclusion: Granulosa cells from human small antral follicles differentiated during culture into cells with a gene expression profile similar to GC from preovulatory follicles.