Culture environment modulates maturation and metabolism of human oocytes.

BACKGROUND The clinical use of oocytes matured in vitro for IVF is increasing, but little is known about the effect of culture conditions on oocyte maturation. METHODS Denuded immature oocytes identified following superovulation prior to ICSI were individually matured in one of two commercial media: tissue culture medium (TCM) 199 or modified Eagle medium with Earle's modified salts (MEME). During maturation, depletion of pyruvate and accumulation of lactate in the culture medium were non-invasively measured. RESULTS Maturing oocytes took up pyruvate (20-30 pmol/oocyte/h) and produced lactate (2-10 pmol/oocyte/h). Oocytes matured faster in MEME, with significantly more oocytes reaching metaphase II by 24 h after oocyte retrieval compared with TCM 199 (P = 0.03). The oocytes that matured more quickly in MEME had significantly lower lactate production than oocytes that matured more slowly (P = 0.02). In TCM 199, pyruvate uptake by rapidly maturing oocytes was lower than by slowly maturing oocytes (P = 0.05). During the second incubation, from 24 to 48 h post-oocyte retrieval, pyruvate uptake in MEME was 30% lower than in TCM 199 (P = 0.007). Pyruvate uptake and lactate production differed depending on the stage of nuclear maturation: pyruvate uptake and lactate production were greater during germinal vesicle breakdown than during polar body extrusion in MEME (P < 0.05). CONCLUSIONS We have shown that: (i). pyruvate is a major energy source during oocyte maturation; (ii). the composition of the culture medium can affect the rate of maturation; and (iii). the culture medium and stage of nuclear maturation can affect pyruvate uptake and lactate production.

[1]  Usa Serono Symposia,et al.  Preimplantation Embryo Development , 1993, Serono Symposia, USA Norwell, Massachusetts.

[2]  P. D. De Sousa,et al.  Impact of Bovine Oocyte Maturation Media on Oocyte Transcript Levels, Blastocyst Development, Cell Number, and Apoptosis1 , 2000, Biology of reproduction.

[3]  D. Levran,et al.  Incubation with sperm enhances in vitro maturation of the oocyte from the germinal vesicle to the M2 stage. , 1997, Fertility and sterility.

[4]  R. Edwards,et al.  Maturation in vitro of Mouse, Sheep, Cow, Pig, Rhesus Monkey and Human Ovarian Oocytes , 1965, Nature.

[5]  D. Gardner,et al.  Temporal and differential effects of amino acids on bovine embryo development in culture. , 1999, Biology of reproduction.

[6]  R. Edwards,et al.  Uptake of pyruvate by early human embryos determined by a non-invasive technique. , 1986, Human reproduction.

[7]  M. Bartolomei,et al.  Differential Effects of Culture on Imprinted H19 Expression in the Preimplantation Mouse Embryo1 , 2000, Biology of reproduction.

[8]  Z. Nagy,et al.  Maturation of human cumulus-free germinal vesicle-stage oocytes to metaphase II by coculture with monolayer Vero cells. , 1995, Human reproduction.

[9]  H. Leese,et al.  Non-invasive measurement of pyruvate and glucose uptake and lactate production by sigle human preimplantation embryos , 1990 .

[10]  S. M. Downs,et al.  The participation of energy substrates in the control of meiotic maturation in murine oocytes. , 1994, Developmental biology.

[11]  A. Trounson,et al.  Maturation of human oocytes in vitro and their developmental competence. , 2001, Reproduction.

[12]  C Wrenzycki,et al.  Alterations of expression of developmentally important genes in preimplantation bovine embryos by in vitro culture conditions: implications for subsequent development. , 2000, Theriogenology.

[13]  H. Leese,et al.  Non-invasive measurement of pyruvate and glucose uptake and lactate production by single human preimplantation embryos. , 1990, Human reproduction.

[14]  B. Pukazhenthi,et al.  Oocyte metabolism predicts the development of cat embryos to blastocyst in vitro , 2000, Molecular reproduction and development.

[15]  Z. Nagy,et al.  Fertilization and early embryology: Maturation of human cumulus-free germinal vesiclestage oocytes to metaphase II by coculture with monolayer Vero cells , 1995 .

[16]  I Wilmut,et al.  Large offspring syndrome in cattle and sheep. , 1998, Reviews of reproduction.

[17]  W. Reik,et al.  Culture of preimplantation embryos and its long-term effects on gene expression and phenotype. , 2001, Human reproduction update.

[18]  S. Franks,et al.  Fertilization and early embryology: Normal development and metabolic activity of preimplantation embryos in vitro from patients with polycystic ovaries , 1995 .

[19]  H. Leese,et al.  Production of pyruvate by isolated mouse cumulus cells. , 1985, The Journal of experimental zoology.

[20]  S. Emiliani,et al.  Amino acids promote human blastocyst development in vitro. , 2001, Human reproduction.

[21]  J. Biggers,et al.  The pattern of energy metabolism in the mouse oöcyte and zygote. , 1967, Proceedings of the National Academy of Sciences of the United States of America.

[22]  B. K. Kim,et al.  In vitro maturation, fertilization, and development of human germinal vesicle oocytes collected from stimulated cycles. , 2000, Fertility and sterility.

[23]  D. Gardner,et al.  Metabolism of glucose, pyruvate, and glutamine during the maturation of oocytes derived from pre‐pubertal and adult cows , 1999, Molecular reproduction and development.

[24]  Y. Englert,et al.  Effects of taurine on human embryo development in vitro. , 1999, Human reproduction.

[25]  B. Bavister,et al.  Effect of oocyte maturation medium on in vitro development of in vitro fertilized bovine embryos , 1992, Molecular reproduction and development.

[26]  S. Downs,et al.  The influence of glucose, cumulus cells, and metabolic coupling on ATP levels and meiotic control in the isolated mouse oocyte. , 1995, Developmental biology.

[27]  S. McKiernan,et al.  Regulation of Hamster Embryo Development In Vitro by Amino Acids , 1993 .

[28]  G. Wright,et al.  Birth after cryopreservation of immature oocytes with subsequent in vitro maturation. , 1998, Fertility and sterility.

[29]  R. Schultz,et al.  Preimplantation development of mouse embryos in KSOM: Augmentation by amino acids and analysis of gene expression , 1995, Molecular reproduction and development.

[30]  H. Leese,et al.  Metabolism of pyruvate by the early human embryo. , 1998, Biology of reproduction.

[31]  D. Rieger,et al.  Changes in the metabolism of glucose, pyruvate, glutamine and glycine during maturation of cattle oocytes in vitro. , 1994, Journal of reproduction and fertility.

[32]  J. Conaghan,et al.  Delaying transfer to the third day post-insemination, to select non-arrested embryos, increases development to the fetal heart stage. , 1995, Human reproduction.

[33]  J. G. Thompson,et al.  Human assisted conception: a cautionary tale. Lessons from domestic animals. , 1998, Human reproduction.

[34]  H. Leese,et al.  Fertilization and early embryology: Comparison of pyruvate uptake by embryos derived from conception and non-conception natural cycles , 1994 .

[35]  G. Pincus,et al.  THE COMPARATIVE BEHAVIOR OF MAMMALIAN EGGS IN VIVO AND IN VITRO , 1935, The Journal of experimental medicine.

[36]  S. Downs,et al.  Precursors of the purine backbone augment the inhibitory action of hypoxanthine and dibutyryl cAMP on mouse oocyte maturation. , 1998, The Journal of experimental zoology.

[37]  J. Leeton,et al.  Environment of the preimplantation human embryo in vivo: metabolite analysis of oviduct and uterine fluids and metabolism of cumulus cells. , 1996, Fertility and sterility.

[38]  Y. Englert,et al.  Noninvasive assessment of glucose and pyruvate uptake by human embryos after intracytoplasmic sperm injection and during the formation of pronuclei. , 2000, Fertility and sterility.

[39]  A. Schroeder,et al.  Culture media for mouse oocyte maturation affect subsequent embryonic development , 1990, Molecular reproduction and development.

[40]  T. Rose-Hellekant,et al.  Energy substrates and amino acids provided during in vitro maturation of bovine oocytes alter acquisition of developmental competence , 1998, Zygote.

[41]  D. Whittingham,et al.  Metabolism of specifically labelled pyruvate by mouse embryos during culture from the two-cell stage to the blastocyst. , 1970, Australian journal of biological sciences.

[42]  R. Winston,et al.  Glutamine improves human preimplantation development in vitro. , 1998, Fertility and sterility.

[43]  C. Fagbohun,et al.  Requirement for glucose in ligand-stimulated meiotic maturation of cumulus cell-enclosed mouse oocytes. , 1992, Journal of reproduction and fertility.

[44]  A. Trounson,et al.  Mitochondrial morphology during preimplantational human embryogenesis. , 2000, Human reproduction.

[45]  J. Robinson,et al.  Developmental consequences of embryo and cell manipulation in mice and farm animals. , 2001, Reproduction.

[46]  H. Leese,et al.  Activity of enzymes of energy metabolism in single human preimplantation embryos. , 1993, Journal of reproduction and fertility.

[47]  B Dale,et al.  Mitochondrial aggregation patterns and activity in human oocytes and preimplantation embryos. , 2001, Human reproduction.

[48]  D. Gardner,et al.  Amino acids and ammonium regulate mouse embryo development in culture. , 1993, Biology of reproduction.

[49]  Z. Nagy,et al.  Pregnancy and birth after intracytoplasmic sperm injection of in vitro matured germinal-vesicle stage oocytes: case report**Supported by grants by the Belgian Fund for Medical Research, Brussels, Belgium , 1996 .

[50]  J. Eppig,et al.  Hypoxanthine and adenosine in murine ovarian follicular fluid: concentrations and activity in maintaining oocyte meiotic arrest. , 1985, Biology of reproduction.

[51]  P. Sutter,et al.  In-vitro maturation of human germinal vesicle stage oocytes: role of cumulus cells and epidermal growth factor in the culture medium. , 1998, Human reproduction.

[52]  H. Niemann,et al.  Energy Metabolism in Preimplantation Bovine Embryos Derived In Vitro or In Vivo1 , 2000, Biology of reproduction.

[53]  A. Handyside,et al.  Non-invasive measurement of glucose and pyruvate uptake by individual human oocytes and preimplantation embryos. , 1989, Human reproduction.

[54]  S. Downs,et al.  Hypoxanthine is the principal inhibitor of murine oocyte maturation in a low molecular weight fraction of porcine follicular fluid. , 1985, Proceedings of the National Academy of Sciences of the United States of America.