Abstract
Purpose: The objectives of these in vitro maturation studies in primate cumulus-oocyte complexes (COCs) were to evaluate the effect of a reduced-oxygen environment and to compare medium with a high-glucose concentration to medium with pyruvate but no glucose.
Methods: COCs were retrieved from squirrel monkeys stimulated with 1 mg of follicle-stimulating hormone (FSH) for 4–6 days. Experiment 1 examined maturation after 48 hr in 5% O 2 /5% CO 2 /90% N 2 compared with 5% CO 2 /air. The medium was CMRL-1066 containing moderate glucose (5.5 mM) supplemented with 1 mM glutamine, 0.33 mM pyruvate, 0.075 IU/ml human FSH, 5 IU/ml human chorionic gonadotropin, 75 U penicillin G/ml, and 20% fetal bovine serum. Experiment 2 in 5% CO 2 /air, compared P-l medium (pyruvate and lactate but no glucose) to Waymouth's medium (27.5 mM glucose), both with identical supplements.
Results: Only 3 (8%) of 37 COCs matured in 5% O 2 , while 39 (49%) of 80 matured in ambient O 2. Fourteen (22%) of 64 complexes matured in P-1 medium, compared to 47 (49%) of 96 meiosis II oocytes in Waymouth's medium (P < 0.05).
Conclusions: These are the first primate studies showing detrimental effects of reduced-oxygen culture on in vitro maturation. Additionally, maturation was enhanced with complex high-glucose medium suggesting that the predominant metabolism is aerobic glycolysis.
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REFERENCES
Eppig JJ: Factors controlling mammalian oocyte maturation. In The Primate Ovary, RL Stouffer (ed), New York, Plenum, 1987, pp 77-90
Alberts B, Dray D, Lewis J, Raff M, Roberts K, Watson JD: The Molecular Biology of the Cell, New York, Garland, 1994
Sorensen RA, Wassarman PM: Relationship between growth and meiotic maturation of the mouse oocyte. Dev Biol 1976;50:531-536
Motlik J, Crozet N, Fulka J: Meiotic competence in vitro of pig oocytes isolated form early antral follicles. J Reprod Fertil 1984;72:323-328
Schramm RD, Tennier MT, Boatman DE, Bavister BD: Chromatin configurations and meiotic competence of oocytes are related ot follicular diameter in ninstimulated rhesus monkeys. Biol Reprod 1993;48:346-356
Gilchrist RB, Nayudu PL, Nowshari MA, Hodges JK: Meiotic competence of marmoset monkey oocytes is related to follicle size and oocyte-somatic cell associations. Biol Reprod 1995;52:1234-1243
Fulka J, Motlik J, Fulka J, Jilek F: Effect of cyclohexamide on nuclear maturation of pig and mouse oocytes. J Reprod Fertil 1986;77:281-285
Bar-Ami S, Seibel MM: Oocyte development and meiosis in humans. In Infertility, MM Seibel (ed). Stamford CT, Appleton and Lange, 1997, pp 61-110
Dekel N, Galiani D, Sherizly I: Dissociation between the inhibitory and the stimulatory action of cAMP on maturation of rat oocytes. Mol Cell Endocrinol 1988;56:115-121
Mattioli M, Galeati G, Bacci ML, Barboni B: Changes in maturation promoting activity in the cytoplasm of pig oocytes throughout maturation. Mol Reprod Dev 1991;30:119-123
Long CR, Damiani P, Pinto-Correia C, MacLean RA, Duby RT, Robl JM: Morphology and subsequent development in culture of bovine oocytes matured in vitro under various conditions of fertilization. J Reprod Fertil 1994;102:361-369
Biggers JD: Metabolism of the Oocyte. In Oogenesis, JD Biggers, AW Schultz (eds). Baltimore, University Park Press, 1972, pp 241-251
Bornslaeger EA, Schultz RM: Regulation of mouse oocyte maturation: Effect of elevating cumulus cell cAMP on oocyte cAMP levels. Biol Reprod 1985;33:698-704
Anderson E, Albertini DF: Gap junctions between the oocyte and companion follicle cells in the mammalian ovary. J Cell Biol 1976;71:680-686
Schramm RD, Bavister BD: Follicle-stimulating hormone priming of rhesus monkeys enhances meiotic and developmental competence of oocytes matured in vitro. Biol Reprod 1994;51:904-912
Yeoman RR, Helvacioglu A, Williams LE, Aksel S, Abee CR: Restoration of oocyte maturational competency during the nonbreeding season with follicle-stimulating hormone stimulation in squirrel monkeys (Saimiri boliviensis boliviensis). Biol Reprod 1994;50:329-335
Alak BM, Wolf DP: Rhesus monkey oocyte maturation and fertilization in vitro: Roles of the menstrual cycle phase and of exogenous gonadotropins. Biol Reprod 1994;51:879-887
Downs SM, Mastropolo AM: Culture conditions affect meiotic regulation in cumulus cell-enclosed mouse oocytes. Mol Reprod Dev 1997;46:551-566
Scott L, Whittingham DG: Influence of genetic background and media components on the development of mouse embryos in vitro. Mol Reprod Dev 1996;43:336-346
Eppig JJ, Wigglesworth K: Factors affecting the developmental competence of mouse oocytes grown in vitro: Oxygen concentration. Mol Reprod Dev 1995;42:447-456
Liu Z, Foote RH: Development of bovine embryos in KSOM with added superoxide dismutase and taurine and with five and twenty percent oxygen. Biol Reprod 1995;53:786-790
Downs SM, Houghton FD, Humpherson PG, Leese HJ: Substrate utilization and maturation of cumulus cell-enclosed mouse oocytes: Evidence that pyruvate oxidation does not mediate meiotic induction. J Reprod Fertil 1997;110:1-10
Gardner DK, Leese HJ: Concentrations of nutrients in mouse oviduct fluid and their effects on embryo development and metabolism in vitro. J Reprod Fertil 1990;88:361-368
Gosden RG, Byatt-Smith JG: Oxygen concentration gradient across the ovarian follicular epithelium: model, predictions and implications. Hum Reprod 1986;1:65-68
Knudsen JF, Litkowshi LJ, Wilson TL, Guthrie HD, Batta SK: Concentrations of hydrogen ions, oxygen, carbon dioxide and bicarbonate in porcine follicular fluid. J Endocrinol 1978;79:249-250
Zeilmaker GH, Jlsmann WC, Wensinck F, Verhamme C: Oxygen triggered oocyte maturation in vitro and lactate utilization by mouse oocytes and zygotes. J Reprod Fertil 1972;9:151-152
Fisher B, Bavister BD: Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits. J Reprod Fertil 1993;99:673-679
Barbehenn EK, Wales RG, Lowry OH: The explanation for the blockade of glycolysis in early mouse embryos. Proc Natl Acad Sci USA 1974;71:1056-1060
Gwatkin RBL, Haidri AA: Oxygen requirements for the maturation of hamster oocytes. J Reprod Fertil 1974;37:127-129
Smitz J, Cortvrindt R, Van Steirteghem AC: Normal oxygen atmosphere is essential for the solitary long-term culture of early preantral mouse follicles. Mol Reprod Dev 1996;45:466-475
Volarcik K, Sheean L, Goldfarb J, Woods L, Abdul-karim FW, Hunt P: The meiotic competence of in-vitro matured human oocytes is influenced by donor age: evidence that folliculogensis is compromised in the reproductively aged ovary. Hum Reprod 1998;13:154-160
Abeydeera LR, Wang W, Prather RS, Day BN: Maturation in vitro of pig oocytes in protein-free culture media: Fertilization and subsequent embryo development in vitro. Biol Reprod 1998;58:1316-1320
Van de Stadt JJM, Schroeder AC, Eppig JJ: Culture media for mouse oocyte maturation affect subsequent embryonic development. Mol Reprod Dev 1990;25:164-171
Leese HJ, Lenton EA: Gluconse and lactate in human follicular fluid: Concentrations and interrelationships. Hum Reprod 1990;5:915-919
Gardner DK, Lane M, Calderon I, Leeton J: Environment of the preimplantation human embryo in vivo: Metabolite analysis of oviduct and uterine fluids and metabolism of cumulus cells. Fertil Steril 1996;65:349-353
Carroll J, Whittingham DG, Wood MJ: Effect of gonadotropin environment on growth and development of isolated mouse primary ovarian follicles. J Reprod Fertil 1991;93:71-77
Kito S, Bavister BD: Gonadotropins, serum, and amino acids alter nuclear maturation, cumulus expansion, and oocyte morphology in hamster cumulus-oocyte complexes in vitro. Biol Reprod 1997;56:1281-1289
Boland NI, Humpherson PG, Leese HJ, Gosden RG: The effect of glucose metabolism on murine follicle development and steroidogenesis in vitro. Hum Reprod 1994;9:617-623
Watson AJ, Watson PH, Warnes D, Walker SK, Armstrong DT, Seamark: Preimplantation development of in vitro-matured and in vitro-fertilized ovine zygotes: Comparison between coculture on oviduct epithelial cell monolayers and culture under low oxygen atmosphere. Biol Reprod 1994;50:715-724
Meister A: Glutathione deficiency produced by inhibition of its synthesis and its reversal; Application in research and therapy. Pharmacol Therap 1991;51:155-194
Yoshida M, Kshigaka K, Nagai T, Chikyu M, Pursel VG: Glutathione concentration during maturation and after fertilization in pig oocytes: Relevance to the ability of oocytes to form male pronucleus. Biol Reprod 1993; 49:89-94
Kito S, Bavister BD: Male pronuclear formation and early embryonic development of hamster oocytes matured in vitro with gonadotropins, amino acids and cysteamine. J Reprod Fertil 1997;110:35-40
Warburg O: The Metabolism of Tumours. London, Arnold Constable, 1930
Brand KA, Hermfisse U: Aerobic glycolysis by proliferating cells: A protective strategy against reactive oxygen species. FASEB J 1997;11:388-395
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Yeoman, R.R., Williams, L.E. & Abee, C.R. Low Oxygen Inhibits but Complex High-Glucose Medium Facilitates In Vitro Maturation of Squirrel Monkey Oocyte–Granulosa Cell Complexes. J Assist Reprod Genet 16, 102–107 (1999). https://doi.org/10.1023/A:1022525108241
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DOI: https://doi.org/10.1023/A:1022525108241