Table 1.
Description of maturation media, including treatment abbreviation, rate of supplementation, hormone concentration, and size of follicle from which the fluid was collected.
Table 2.
Area of cumulus oocyte complexes (COCs; μm2) at 0 and 19 h of the maturation period. Cumulus oocyte complexes (n=480) were matured in one of two control or ten follicular fluid-containing media (75% follicular fluid supplementation). Follicular fluid originated from large or small follicles and was either untreated, charcoal stripped, or charcoal stripped and supplemented with estradiol and/or progesterone. Data presented as least squares mean ± SEM (x100000).
Fig 1.
Effect of follicular fluid and steroids during the maturation period on cumulus cell expansion (percentage of area increase).
Cumulus oocyte complexes (n=480) were matured in oocyte maturation medium supplemented with 0% (cOMM, eOMM) or various combinations of follicular fluid and steroids throughout the maturation period. cOMM= control oocyte maturation medium; eOMM= experimental OMM (contains charcoal stripped (cs) fetal bovine serum, no FSH, no estradiol and no EGF); LFF75= 75% untreated large follicular fluid; SFF75= 75% untreated small follicular fluid; csLFF75= 75% charcoal stripped large follicular fluid; csSFF75= 75% charcoal stripped small follicular fluid; E2= estradiol (23 ng/ml for csSFF and 37 ng/ml for csLFF treatments); P4= progesterone (140 ng/ml for csSFF and 160 ng/ml for csLFF treatments). Different letters above the bars indicate significant difference between treatments (P≤0.04).
Fig 2.
Representative image of oocyte maturation in response to exposure to follicular fluid.
(A-L) Cumulus oocyte complexes (COCs) were matured for 19 h in the presence or absence of 75% follicular fluid as an additive to the oocyte maturation medium; cOMM= control oocyte maturation medium; eOMM= experimental OMM (contains charcoal stripped (cs) fetal bovine serum, no FSH, no estradiol and no EGF); LFF75= 75% untreated large follicular fluid; SFF75= 75% untreated small follicular fluid; csLFF75= 75% charcoal stripped large follicular fluid; csSFF75= 75% charcoal stripped small follicular fluid; E2= estradiol (23 ng/ml for csSFF and 37 ng/ml for csLFF treatment); P4= progesterone (140 ng/ml for csSFF and 160 ng/ml for csLFF treatment). (M) Representative COCs at the time of collection and beginning of maturation (time 0, t 0h). Scale bar on figures A-M = 400µm; images captured at 10X. (N) Denuded mature oocyte with the extruded polar body (PB). DNA was labeled with Hoescht 33342 (blue). There were no differences in the nuclear maturation rates as an effect of treatment. Scale bar on panel N = 50µm; image captured at 40X magnification.
Fig 3.
Effect of follicular fluid and steroids during the maturation period on cleavage rates.
Cumulus oocyte complexes (n=4,006) were matured in oocyte maturation medium supplemented with 0% (cOMM, eOMM) or various combinations of follicular fluid and steroids throughout the maturation period. cOMM= control oocyte maturation medium; eOMM= experimental OMM (contains charcoal stripped (cs) fetal bovine serum, no FSH, no estradiol and no EGF); LFF75= 75% untreated large follicular fluid; SFF75= 75% untreated small follicular fluid; csLFF75= 75% charcoal stripped large follicular fluid; csSFF75= 75% charcoal stripped small follicular fluid; E2= estradiol (23 ng/ml for csSFF and 37 ng/ml for csLFF treatments); P4= progesterone (140 ng/ml for csSFF and 160 ng/ml for csLFF treatments). Different letters above the bars indicate significant difference between treatments (P≤0.05).
Fig 4.
Effect of follicular fluid and steroids during the maturation period on blastocyst development.
Cumulus oocyte complexes (n=4,006) were matured in oocyte maturation medium supplemented with 0% (cOMM, eOMM) or various combinations of follicular fluid and steroids throughout the maturation period. cOMM= control oocyte maturation medium; eOMM= experimental OMM (contains charcoal stripped (cs) fetal bovine serum, no FSH, no estradiol and no EGF); LFF75= 75% untreated large follicular fluid; SFF75= 75% untreated small follicular fluid; csLFF75= 75% charcoal stripped large follicular fluid; csSFF75= 75% charcoal stripped small follicular fluid; E2= estradiol (23 ng/ml for csSFF and 37 ng/ml for csLFF treatments); P4= progesterone (140 ng/ml for csSFF and 160 ng/ml for csLFF treatments). Different letters above the bars indicate significant difference between treatments (P≤0.05).