OBJECTIVE: Mammalian follicle cells are the most important somatic cells which help oocytes grow, mature and ovulate and thus are believed to provide oocytes with various functional and structural components. In the present study we have examined whether cumulus or granulosa cells might play a role in establishing the plasma membrane structure of mouse oocytes during meiotic maturation. DESIGN: In particular the differential resistances of mouse oocytes against chymotrypsin treatment were examined following culture with or without cumulus or granulosa cells, or in these cell-conditioned media. RESULTS: When mouse denuded oocytes, freed from their surrounding cumulus cells, were cultured in vitro for 17~18 hr and then treated with 1% chymotrypsin, half of the oocytes underwent degeneration within 37.5 min (t50=37.5+/-7.5 min) after the treatment. In contrast cumulus-enclosed oocytes showed t50=207.0. Similarly, when oocytes were co-cultured with cumulus cells which were not associated with the oocytes but present in the same medium, the t50 of co-cultured oocytes was 177.5+/-13.1 min. Furthermore, when oocytes were cultured in the cumulus cell-conditioned medium, t50 of these oocytes was 190.0+/-10.8 min whereas t50 of the oocytes cultured in M16 alone was 25.5+/-2.9 min. Granulosa cell-conditioned medium also increased the resistance of oocytes against chymotrypsin treatment such that t50 of oocytes cultured in granulosa cell-conditioned medium was 152.5+/-19.0 min while that of oocytes cultured in M16 alone was 70.0+/-8.2 min. To see what molecular components of follicle cell-conditioned medium are involved in the above effects, the granulosa cell-conditioned medium was separated into two factions by using Microcon-10 membrane filter having a 10 kDa cut-off range. When denuded oocytes were cultured in medium containing the retentate, t50 of the oocytes was 70.0+/-10.5 min. In contrast, t50 of the denuded oocytes cultured in medium containing the filtrate was 142.0+/-26.5 min. T50 of denuded oocytes cultured in medium containing both retentate and filtrate was 188.0+/- 13.6 min. However, t50 of denuded oocytes cultured in M16 alone was 70.0 +/-11.0 min and that of oocytes cultured in whole granulosa cell-conditioned medium was 156.0+/-27.9 min. When surface membrane proteins of oocytes were electrophoretically analyzed, no difference was found between the protein profiles of oocytes cultured in M16 alone and of those cultured in the filtrate. CONCLUSIONS: Based upon these results, it is concluded that mouse follicle cells secrete a factor(s) which enhance the resistance of mouse oocytes against a proteolytic enzyme treatment. The factor appears to be a small molecules having a molecular weight less than 10 kDa.
Animals ; Cell Membrane ; Chymotrypsin* ; Cumulus Cells ; Female ; Granulosa Cells ; Membrane Proteins ; Membranes ; Mice* ; Molecular Weight ; Oocytes*

No abstract available.
Female ; Granulosa Cell Tumor* ; Granulosa Cells* ; Ovary*

No abstract available.
Female ; Granulosa Cell Tumor ; Granulosa Cells

No abstract available.
Female ; Granulosa Cell Tumor* ; Granulosa Cells* ; Humans ; Infant*

No abstract available.
Female ; Granulosa Cell Tumor ; Granulosa Cells ; Neoplasm Metastasis ; Ovary

Granulosa cell tumors of the ovary are rare, and account for 2 to 3% of ovarian tumors. Granulosa cell tumors are discovered often in perimenopausal or postmenopausal women but 10 to 26% are found in the reproductive age group. The tumors associated with pregnancy are infrequent. In this study, we present a case of granulosa cell tumor of left ovary at pregnancy.
Female ; Granulosa Cell Tumor* ; Granulosa Cells* ; Humans ; Ovary ; Pregnancy*

In this study, in order to further understanding of function of Mullerian inhibiting substance (MIS) and the ontogeny of the production profile of biologically active MIS and MIS type II receptor (MISR II), the patterns of their localization according to the follicular development in 21 ovarian specimens from women in reproductive age were studied by immunohistochemical staining. The flattened granulosa cells in primordial follicles failed to stain for MIS and MISR II, but the first staining was detected in the cuboidal granulosa cells in primary follicles. MIS and MISR II were detected specifically and exclusively in the cytoplasm of granulosa cells. The granulosa cells of both single and multiple layered growing preantral follicles showed strong specific staining for MIS and MISR II. Among the growing follicles, large follicle stained more intensely than small one. Within the multiple layers of granulosa cells, the innermost cells, closer to the oocyte, stained more intensely for MIS than those near the basement membrane, but MISR II was evenly distributed. In antral follicles, expression of the MIS was only seen in the granulosa cells, but MISR II was seen in the granulosa cells and theca cells. In large antral follicles, cumulus cells and periantral granulosa cells stained more intensely for MIS than those in the periphery. MIS staining waned in the mature follicles just before ovulation and could not be found in atretic follicles, corpus luteum, and corpus albicans. The expression levels of MISR II in mature follicles was lower than those in growing follicles and were even further reduced, but still detectable, in corpus luteum. There was a decreased level of MISR II expression when follicles become atretic and eventually lost from atretic follicles. The MIS and MISR II staining were not found in primordial follicles, oocytes, interstitial cells, ovarian epithelium, and corpus albicans. It is concluded that actions of MIS via MISR II are autocrine and paracrine in nature. The pattern of MIS and MISR II expression according to the menstrual cycles and development suggest that MIS may act as an intraovarian regulator of follicle maturation, selection and ovulation during the adult reproductive cycle.
Adult ; Anti-Mullerian Hormone* ; Basement Membrane ; Corpus Luteum ; Cumulus Cells ; Cytoplasm ; Epithelium ; Female ; Granulosa Cells ; Humans* ; Menstrual Cycle* ; Oocytes ; Ovarian Follicle ; Ovary* ; Ovulation ; Theca Cells

Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and preimplantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1% chymotrypsin, half of them remained still alive even after 240 min (t50>240.0). In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min (t50= 65.0+/-13.2 min) of the same treatment. To see if the effects were due to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited t50 = 190.0 +/-10.8 min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited t50 = 183.3+/-19.1 min while t50 of control group oocytes was 60.0+/-6.8 min. Oocytes matured in vero cell conditioned medium exhibited t50 = 196.7+/-8.8 min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of t50 = 80.0+/-8.4 min which was not statistically different from the control value of t50 = 48.0+/-13.2 min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.
Animals ; Chymotrypsin* ; Culture Media, Conditioned* ; Cumulus Cells ; Eggs ; Female ; Fertilization in Vitro ; Granulosa Cells ; Hand ; Humans ; Membranes ; Mice* ; Oocytes* ; Ovum ; Vero Cells

Mullerian inhibiting substance(MIS) has been known as a non-steroidal testicular Sertolicell product responsible for the regression of Mullerian duct in male embryos. More recently MIS was also found to be present in an bioactive form in the bovine and rat ovaries but the function of MIS in the ovary has not been fully delineated. In this study, in order to understand its function in the ovary the ontogeny of the production profile of MIS and the pattern of its localization in ovaries from adult normal cycling women were studied by immunohistochemical staining using the rabbit polyclonal antibody against human recombinant MIS that almost completely blocks its biological activity. MIS was detected specifically and exclusively in the cytoplasm of granulosa cells. The flattened granulosa cells in primordial follicles failed to stain for MIS, but the cuboidal cells of growing follicles stained intensely. The granulosa cells of both single and multiple layered growing follicles showed strong specific staining for MIS. Within the multiple layers of granulosa cells, closer to the oocyte, stained more intensely than those near the basement membrane. Similarly, in antral follicles, cumulus cells and periantral granulosa cells stained more intensely than those in the periphery. MIS staining waned in the mature follicles just before ovulation and could not be found in atretic follicles, corpus albicans. In conclusion, this specific localization suggest that MIS may act as an intraovarian regulator of follicular development and oocyte maturation during the adult reproductive cycle.
Adult ; Animals ; Anti-Mullerian Hormone* ; Basement Membrane ; Cumulus Cells ; Cytoplasm ; Embryonic Structures ; Female ; Granulosa Cells ; Humans* ; Male ; Oocytes ; Ovarian Follicle ; Ovary* ; Ovulation ; Rats

OBJECTIVE: This study was performed to explore the possibility that each oocyte and its surrounding cumulus cells might have different genetic expression patterns that could affect human reproduction. METHODS: Differential gene expression analysis was performed for 10 clusters of cumulus cells obtained from 10 cumulus-oocyte complexes from 10 patients. Same procedures related to oocyte maturation, microinjection, and microarray analyses were performed for each group of cumulus cells. Two differential gene expression analyses were performed: one for the outcome of clinical pregnancy and one for the outcome of live birth. RESULTS: Significant genes resulting from these analyses were selected and the top 20 affected pathways in each group were analyzed. Circadian entrainment is determined to be the most affected pathway for clinical pregnancy, and proteoglycans in cancer pathway is the most affected pathway for live birth. Circadian entrainment is also amongst the 12 pathways that are found to be in top 20 affected pathways for both outcomes, and has both lowest p-value and highest number of times found count. CONCLUSION: Although further confirmatory studies are necessary, findings of this study suggest that these pathways, especially circadian entrainment in cumulus cells, may be essential for embryo development and pregnancy.
Circadian Clocks ; Cumulus Cells ; Embryonic Development ; Female ; Gene Expression ; Granulosa Cells ; Humans ; Infertility ; Live Birth ; Microarray Analysis ; Microinjections ; Oocytes ; Ovarian Follicle ; Pregnancy ; Proteoglycans ; Reproduction ; Reproductive Techniques, Assisted