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Cumulus cells steroidogenesis is influenced by the degree of oocyte maturation.

Lucidi P, Bernabò N, Turriani M, Barboni B, Mattioli M - Reprod. Biol. Endocrinol. (2003)

Bottom Line: The secretion of the factor(s) in conditioned media was then recorded by evaluating the ability of the spent media to direct granulosa cells (GC) steroidogenesis.Monolayers obtained by growing GC surrounding the oocytes for five days represent a tool, which is practical, stable and available in most laboratories; by using this bioassay, we detected the antiluteal effect of immature oocytes, and for the first time, demonstrated that properly matured germ cells are able to direct cumulus cells steroidogenesis by inhibiting E2 production (P < 0.01).Nevertheless, only fully competent oocytes were able to suppress estrogens production, while those cultured under unfavourable conditions were unable to exert any inhibitory effect on the functions of cumulus cells (P < 0.01).

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Scienze Biomediche Comparate, Facoltà di Medicina Veterinaria, Università degli Studi di Teramo, Piazza Aldo Moro 45, Italia. lucidi@unite.it

ABSTRACT

Background: The possibility to predict the ability of a germ cell to properly sustain embryo development in vitro or in vivo as early as possible is undoubtedly the main problem of reproductive technologies. To date, only the achievement of nuclear maturation and cumulus expansion is feasible, as all the studies on cytoplasmic maturation are too invasive and have been complicated by the death of the cells analyzed. The authors studied the possibility to test the cytoplasmic quality of pig oocytes by evaluating their ability to produce steroidogenesis enabling factor(s). To this aim, oocytes matured under different culture conditions that allowed to obtain gradable level of cytoplasmic maturation, were used to produce conditioned media (OCM). The secretion of the factor(s) in conditioned media was then recorded by evaluating the ability of the spent media to direct granulosa cells (GC) steroidogenesis.

Methods: In order to obtain germ cells characterized by a different degree of developmental competence, selected pig oocytes from prepubertal gilts ovaries were cultured under different IVM protocols; part of the matured oocytes were used to produce OCM, while those remaining were submitted to in vitro fertilization trials to confirm their ability to sustain male pronuclear decondensation. The OCM collected were finally used on cumulus cells grown as monolayers for 5 days. The demonstration that oocytes secreted factor(s) can influence GC steroidogenesis in the pig was confirmed in our lab by studying E2 and P4 production by cumulus cells monolayers using a radioimmunoassay technique.

Results: Monolayers obtained by growing GC surrounding the oocytes for five days represent a tool, which is practical, stable and available in most laboratories; by using this bioassay, we detected the antiluteal effect of immature oocytes, and for the first time, demonstrated that properly matured germ cells are able to direct cumulus cells steroidogenesis by inhibiting E2 production (P < 0.01). Nevertheless, only fully competent oocytes were able to suppress estrogens production, while those cultured under unfavourable conditions were unable to exert any inhibitory effect on the functions of cumulus cells (P < 0.01).

Conclusion: These results demonstrated that good quality oocytes can be easily selected on the basis of their ability to affect granulosa cell steroidogenesis thus reducing failures in reproductive technologies due to the transfer of fertilized oocytes with a scarce ability to sustain embryo development.

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Related in: MedlinePlus

Challenge of cumulus cell monolayers with conditioned media from pig oocytes cultured in vitro under different, unfavorable conditions, namely COCs without hormonal support (M-H), and matured denuded oocytes (MD), compared to immature (GV) and properly matured cells (M+H). Histograms represent the mean E2 production ± sd. Control wells were treated as in fig 2 and 3. * P < 0.01 (Student t-test).
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Figure 4: Challenge of cumulus cell monolayers with conditioned media from pig oocytes cultured in vitro under different, unfavorable conditions, namely COCs without hormonal support (M-H), and matured denuded oocytes (MD), compared to immature (GV) and properly matured cells (M+H). Histograms represent the mean E2 production ± sd. Control wells were treated as in fig 2 and 3. * P < 0.01 (Student t-test).

Mentions: In order to know whether attaining an adequate cytoplasmic maturation was important for the production of soluble factor(s) by the oocytes, the cumulus cell monolayers were challenged with conditioned media obtained from MII oocytes. The MII oocytes belonged to the classes of cells that had matured in poor conditions, i.e. matured denuded oocytes (MD), or cells cultured as COCs without hormone support (M-H), or with a complete cell and gonadotropin support (M+H). The results of this challenge (figure 4) demonstrate that oocytes matured in the absence of cellular or hormonal support were unable to inhibit E2 production by cumulus cell monolayers while they confirm (P < 0.01) the inhibitory role of media conditioned with M+H oocytes (600.57 ± 287.9 pg/104 cells/mL vs 1730.33 ± 205.4 pg/104 cells/mL for FSH treated group).


Cumulus cells steroidogenesis is influenced by the degree of oocyte maturation.

Lucidi P, Bernabò N, Turriani M, Barboni B, Mattioli M - Reprod. Biol. Endocrinol. (2003)

Challenge of cumulus cell monolayers with conditioned media from pig oocytes cultured in vitro under different, unfavorable conditions, namely COCs without hormonal support (M-H), and matured denuded oocytes (MD), compared to immature (GV) and properly matured cells (M+H). Histograms represent the mean E2 production ± sd. Control wells were treated as in fig 2 and 3. * P < 0.01 (Student t-test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC161805&req=5

Figure 4: Challenge of cumulus cell monolayers with conditioned media from pig oocytes cultured in vitro under different, unfavorable conditions, namely COCs without hormonal support (M-H), and matured denuded oocytes (MD), compared to immature (GV) and properly matured cells (M+H). Histograms represent the mean E2 production ± sd. Control wells were treated as in fig 2 and 3. * P < 0.01 (Student t-test).
Mentions: In order to know whether attaining an adequate cytoplasmic maturation was important for the production of soluble factor(s) by the oocytes, the cumulus cell monolayers were challenged with conditioned media obtained from MII oocytes. The MII oocytes belonged to the classes of cells that had matured in poor conditions, i.e. matured denuded oocytes (MD), or cells cultured as COCs without hormone support (M-H), or with a complete cell and gonadotropin support (M+H). The results of this challenge (figure 4) demonstrate that oocytes matured in the absence of cellular or hormonal support were unable to inhibit E2 production by cumulus cell monolayers while they confirm (P < 0.01) the inhibitory role of media conditioned with M+H oocytes (600.57 ± 287.9 pg/104 cells/mL vs 1730.33 ± 205.4 pg/104 cells/mL for FSH treated group).

Bottom Line: The secretion of the factor(s) in conditioned media was then recorded by evaluating the ability of the spent media to direct granulosa cells (GC) steroidogenesis.Monolayers obtained by growing GC surrounding the oocytes for five days represent a tool, which is practical, stable and available in most laboratories; by using this bioassay, we detected the antiluteal effect of immature oocytes, and for the first time, demonstrated that properly matured germ cells are able to direct cumulus cells steroidogenesis by inhibiting E2 production (P < 0.01).Nevertheless, only fully competent oocytes were able to suppress estrogens production, while those cultured under unfavourable conditions were unable to exert any inhibitory effect on the functions of cumulus cells (P < 0.01).

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Scienze Biomediche Comparate, Facoltà di Medicina Veterinaria, Università degli Studi di Teramo, Piazza Aldo Moro 45, Italia. lucidi@unite.it

ABSTRACT

Background: The possibility to predict the ability of a germ cell to properly sustain embryo development in vitro or in vivo as early as possible is undoubtedly the main problem of reproductive technologies. To date, only the achievement of nuclear maturation and cumulus expansion is feasible, as all the studies on cytoplasmic maturation are too invasive and have been complicated by the death of the cells analyzed. The authors studied the possibility to test the cytoplasmic quality of pig oocytes by evaluating their ability to produce steroidogenesis enabling factor(s). To this aim, oocytes matured under different culture conditions that allowed to obtain gradable level of cytoplasmic maturation, were used to produce conditioned media (OCM). The secretion of the factor(s) in conditioned media was then recorded by evaluating the ability of the spent media to direct granulosa cells (GC) steroidogenesis.

Methods: In order to obtain germ cells characterized by a different degree of developmental competence, selected pig oocytes from prepubertal gilts ovaries were cultured under different IVM protocols; part of the matured oocytes were used to produce OCM, while those remaining were submitted to in vitro fertilization trials to confirm their ability to sustain male pronuclear decondensation. The OCM collected were finally used on cumulus cells grown as monolayers for 5 days. The demonstration that oocytes secreted factor(s) can influence GC steroidogenesis in the pig was confirmed in our lab by studying E2 and P4 production by cumulus cells monolayers using a radioimmunoassay technique.

Results: Monolayers obtained by growing GC surrounding the oocytes for five days represent a tool, which is practical, stable and available in most laboratories; by using this bioassay, we detected the antiluteal effect of immature oocytes, and for the first time, demonstrated that properly matured germ cells are able to direct cumulus cells steroidogenesis by inhibiting E2 production (P < 0.01). Nevertheless, only fully competent oocytes were able to suppress estrogens production, while those cultured under unfavourable conditions were unable to exert any inhibitory effect on the functions of cumulus cells (P < 0.01).

Conclusion: These results demonstrated that good quality oocytes can be easily selected on the basis of their ability to affect granulosa cell steroidogenesis thus reducing failures in reproductive technologies due to the transfer of fertilized oocytes with a scarce ability to sustain embryo development.

Show MeSH
Related in: MedlinePlus