Intrafollicular Oocyte Maturation Research Articles

Effects of Culture Duration, Follicle Stimulating Hormone (FSH) Type, and Activin A Concentration on In Vitro Growth of Preantral Follicles and Maturation of Intrafollicular Oocytes

Effects of Culture Duration, Follicle Stimulating Hormone (FSH) Type, and Activin A Concentration on In Vitro Growth of Preantral Follicles and Maturation of Intrafollicular Oocytes

Growth of Ovarian Primary Follicles Retrieved from Neonates of Different Ages and Derivation of Mature Oocytes Following In vitro-Culture.

This study was conducted to improve the yield of mature oocytes from in vitro-culture of ovarian primary follicles by optimizing follicle retrieval from neonatal mice of different ages. Primary follicles of 75 to 99 μm in diameter were collected daily from 7- to 14-day-old neonatal mice, and subsequently cultured in α-MEM medium. Number of primary follicles isolated, growth of the follicle during in vitro-culture and maturation of intrafollicular oocytes were monitored. Overall, mean number of preantral follicles per animal was improved from 10.7 to 88.7 as the age of follicle donors was increased from 7 to 14-day-old. Number of primary follicles was increased gradually up to 11-day-old (35.7 follicle per an animal), then reduced to 29 in 14-day-old (p = 0.0013). More follicles retrieved from 10-day-old or 11-day-old females maintained their morphological normality at the end of primary culture than the follicles retrieved from 9-day-old. Of those cultured, primary follicles retrieved from 11-day-old mice yielded largest larger number of early secondary follicles than the follicles retrieved from in the other ages (39 vs. 13 to 29%). More than 3.3-times increase (0.86 to 2.86; p<0.05) in an average number of mature oocytes per animal was observed in the group of 11-day-old, compared with 9-day-old. However, no difference was found in the percentage of primary follicles developing into the pseudoantral stage (21 to 30%; p = 0.5222) and in the percentage of oocytes mucified (32 to 39%; p = 0.5792). In conclusion, a positive correlation between retrieval time and follicle growth was detected, which influences the efficiency to derive mature oocytes by follicle culture.

Effects of -Mercaptoethanol on the Growth of Preantral Follicles and the Maturation of Intrafollicular Oocytes

This study was undertaken to evaluate how β-mercaptoethanol (bME), an exogenous antioxidant, interacts with preantral follicles cultured in vitro. Mouse primary or secondary follicles were cultured in glutathione (GSH)-free or GSH-containing medium supplemented with bME of various concentrations, and the growth of preantral follicles, the maturation of intrafollicular oocytes and preimplantation development after parthenogenesis were monitored. In experiment 1, 0, 25, 50 or 100 μM bME was added to culture medium supplemented with 100 μM GSH or not. When secondary follicles were cultured in GSH-free medium, no significant change in follicle growth was detected after bME addition. However, exposure to bME in the presence of GSH significantly inhibited both follicle growth and oocyte maturation. Such detrimental effect became prominent in primary follicles and bME strongly inhibited follicle growth in the absence of GSH. In conclusion, there are stage-dependent effects of bME on follicle growth and oocyte maturation, and selective use of antioxidants contributes to establishing an efficient follicle culture system.

Establishment of a basic method for manipulating preantral follicles: effects of retrieval method on in vitro growth of preantral follicles and intrafollicular oocytes

The aim of this study was to establish a basic manipulation protocol of preantral follicles for deriving developmentally competent oocytes. Primary, early and late secondary follicles retrieved from the ovaries of 14-day-old F1 (C57BL/6 x DBA2) female mice mechanically or enzymatically were cultured singly and in vitro growth of the follicles and maturation of intrafollicular oocytes were subsequently monitored. A mechanical method retrieved more (p < 0.0001) follicles (339 +/- 48 vs. 202 +/- 28) than an enzymatic method. However, the enzymatic method collected more singly isolated follicles that could be provided for subsequent culture (102 +/- 26 vs. 202 +/- 28). When an enzymatic method was employed, early and late secondary follicles required 9 and 6 days for reaching the maximal incidence of the pseudoantral stage. However, primary follicles were not possible to develop into the pseudoantral stage. The optimal duration of oocyte maturation from the onset of follicle culture was 7 days and 5-7 days for early and late secondary follicles, respectively. A general decrease in oocyte diameter (65.2-65.53 microm vs. 75 microm) and zona thickness (5.41-5.74 microm vs. 7.76 microm) was detected in in vitro-derived compared with in vivo-derived matured oocytes. Pronuclear formation was detected in 86-94% of mature oocytes after parthenogenetic activation and no significant difference was detected among groups. These results showed that preantral follicles retrieved by an enzymatic method underwent step-by-step growth in vitro, which could yield mature oocytes.

Comparison of Follicular and Oocyte Development and Reproductive Hormone Secretion during the Ovulatory Period in Hungarian Native Breed, Mangalica, and Landrace Gilts

Only a very small amount of physiological data is available about the low fertility (mean litter size is 5.7+/-0.8) of Hungarian native breed, Mangalica (M), sows. The aim of the present paper is to reveal the differences in preovulatory follicle development and intrafollicular oocyte maturation between M and Landrace (L) gilts, with special reference to the peri- and postovulatory secretion and peripheral concentrations of estradiol-17beta (E2), progesterone (P4), and luteinizing hormone (LH). The number of preovulatory follicles was 6.8+/-1.4 and 19.6+/-6.6 in M and L gilts, respectively. A lower degree of cumulus expansion and a lower percentage of mature oocytes (TI/M II) was noted in M. Higher LH and E2 peak levels, a longer E2 to LH peak interval, and lower embryo survival was confirmed. Interestingly, despite the lower number of corpora lutea, a higher peripheral blood level of P4 was shown in M than in L gilts. Both diminished follicular development and protracted oocyte maturation may be involved in low fecundity in M, and the present findings may explain these reproductive phenomena.

The effects of low dosages of steroid hormones and carp hypothysial homogenate on carp ( Cyprinus carpio) oocyte maturation in vitro

Using in vitro incubation of carp ovarian follicles, it was shown that low dosages of steroid hormones (0.1–0.5 μg/ml of medium) applied together with carp hypophysial homogenate (c.h.h.) at 100 μg/ml of medium exerted a much more potent effect on intrafollicular oocyte maturation than c.h.h. alone at 100 μg/ml or steroid hormones alone at 0.1–0.5 μg/ml. It is possible that the low dosages of steroid hormones or c.h.h. increased the sensitivity of the follicles to the final gonadotropin stimulation.