Human Reproduction, Vol. 17, No. 5, 1181-1188,
May 2002
© 2002 European Society of Human Reproduction and Embryology
Effects of varying gonadotrophin dose and timing on antrum formation and ovulation efficiency of mouse follicles in vitro
Department of Biological Sciences University of Warwick Coventry CV4 7AL UK
BACKGROUND: This study tested factors affecting mouse follicle growth in vitro, to determine end-points marking follicle function in vitro. METHODS: Pre-antral follicles (mean 137 µm) from B6CBF1 mice were cultured in a substrate-adherent system for 
14 days. FSH (0–1000mIU/ml) day of HCG (1.5 IU/ml days 9–14) protein supplement [fetal calf serum (FCS) (x2) mouse serum (x2) hypogonadal (hpg) mouse serum or human serum albumin (HSA)] were varied. Follicle survival timing of antrum formation incidence of ovulation within 16 24 40 48 h of HCG oocyte growth were assessed. RESULTS: FSH (100 mIU/ml) produced the best antral development (P < 0.001 versus 10 1000 mIU/ml). Antra were observed from day 5. Transient antra formed occasionally in the absence of FSH. By 14 days significant senescence had occurred (P < 0.001) but the proportion of follicles ovulating within 16 h of HCG declined from day 9 onwards indicating this to be a more sensitive marker of follicle responsiveness. Optimal growth occurred in 5% FCS (x2) or hpg mouse serum although fewer follicles ovulated in hpg serum (P < 0.05). No normal growth occurred in normal mouse serum (x2) or HSA. Oocytes grew to full size within 9 days with 100 mIU/ml FSH FCS. CONCLUSIONS: These data provide sensitive end-points for assessing follicle growth in vitro.
Key words: antrum formation/follicle growth in vitro/FSH/in-vitro culture/mouse follicles
1 To whom correspondence should be addressed. E-mail:ghartshorne{at}bio.warwick.ac.uk
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