1,2-propanediol and the type of cryopreservation procedure adversely affect mouse oocyte physiology.

BACKGROUND The aim of this work was to examine the effect of 1,2-propanediol (PrOH) and type of cryopreservation procedure (slow freezing and vitrification) on oocyte physiology. METHODS Intracellular calcium of mouse metaphase II (MII) oocytes was quantified by fluorescence microscopy. The effect of PrOH on cell physiology was further assessed through analysis of zona pellucida hardening and cellular integrity. Protein profiles of cryopreserved oocytes were generated by time-of-flight mass spectrometry (TOF-MS). RESULTS PrOH caused a protracted increase in calcium, which was sufficient to induce zona pellucida hardening and cellular degeneration. Using 'nominally calcium free' media during PrOH exposure significantly reduced the detrimental effects. Proteomic analysis identified numerous up- and down-regulated proteins after slow freezing when compared with control and vitrified oocytes. CONCLUSIONS Using such approaches to assess effects on cellular physiology is fundamental to improving assisted reproduction techniques (ART). This study demonstrates that PrOH causes a significant rise in intracellular calcium. Using calcium-free media significantly reduced the increase in calcium and the associated detrimental physiological effects, suggesting that calcium-free media should be used with PrOH. In addition, analysis of the oocyte proteome following cryopreservation revealed that slow freezing has a significant effect on protein expression. In contrast, vitrification had a minimal impact, indicating that it has a fundamental advantage for the cryopreservation of oocytes.

[1]  E. Porcu,et al.  Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes. , 1997, Fertility and sterility.

[2]  Peter Lipp,et al.  Calcium - a life and death signal , 1998, Nature.

[3]  M. Hirabayashi,et al.  Successful vitrification of pronuclear-stage rabbit zygotes by minimum volume cooling procedure. , 2004, Theriogenology.

[4]  M. Kurokawa,et al.  Intracellular Calcium Oscillations Signal Apoptosis Rather than Activation in In Vitro Aged Mouse Eggs1 , 2002, Biology of reproduction.

[5]  S. Mok,et al.  Protein profiling of cervical cancer by protein-biochips: proteomic scoring to discriminate cervical cancer from normal cervix. , 2004, Cancer letters.

[6]  M. R. Valojerdi,et al.  Developmental potential and ultrastructural injuries of metaphase II (MII) mouse oocytes after slow freezing or vitrification , 2005, Journal of Assisted Reproduction and Genetics.

[7]  D. Gardner,et al.  Proteomic analysis of individual human embryos to identify novel biomarkers of development and viability. , 2006, Fertility and sterility.

[8]  C. Quintans,et al.  Birth of two babies using oocytes that were cryopreserved in a choline-based freezing medium. , 2002, Human reproduction.

[9]  X. Yang,et al.  High Developmental Rates of Vitrified Bovine Oocytes Following Parthenogenetic Activation, In Vitro Fertilization, and Somatic Cell Nuclear Transfer1 , 2000, Biology of reproduction.

[10]  J. Boldt,et al.  Human oocyte cryopreservation as an adjunct to IVF-embryo transfer cycles. , 2003, Human reproduction.

[11]  M. Tucker,et al.  Cryoloop vitrification in assisted reproduction: analysis of survival rates in > 1000 human oocytes after ultra-rapid cooling with polymer augmented cryoprotectants. , 2003, Clinical and experimental obstetrics & gynecology.

[12]  M. Larman,et al.  Calcium-free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes. , 2006, Reproduction.

[13]  I. Jové,et al.  The births of five Spanish babies from cryopreserved donated oocytes. , 2003, Human reproduction.

[14]  P. Halada,et al.  Protein Patterns of Pig Oocytes During In Vitro Maturation1 , 2004, Biology of reproduction.

[15]  K. Kyono,et al.  Successful pregnancy and delivery after transfer of a single blastocyst derived from a vitrified mature human oocyte. , 2005, Fertility and sterility.

[16]  D. Gardner,et al.  Vitrification of mouse oocytes using a nylon loop , 2001, Molecular reproduction and development.

[17]  R. Bast,et al.  Three Biomarkers Identified from Serum Proteomic Analysis for the Detection of Early Stage Ovarian Cancer , 2004, Cancer Research.

[18]  M. Montag,et al.  Developmental rate and ultrastructure of vitrified human pronuclear oocytes after step-wise versus direct rehydration. , 2004, Human reproduction.

[19]  Christopher Chen.  PREGNANCY AFTER HUMAN OOCYTE CRYOPRESERVATION , 1986, The Lancet.

[20]  Andrea Boggio.  Italy enacts new law on medically assisted reproduction. , 2005, Human reproduction.

[21]  H. Sasada,et al.  Vitrification of large quantities of immature bovine oocytes using nylon mesh. , 2001, Cryobiology.

[22]  Shee-Uan Chen,et al.  Cryopreserved oocytes of infertile couples undergoing assisted reproductive technology could be an important source of oocyte donation: a clinical report of successful pregnancies. , 2005, Human reproduction.

[23]  B. Bavister,et al.  Containerless vitrification of mammalian oocytes and embryos , 1999, Nature Biotechnology.

[24]  M. Whitaker,et al.  Sperm-egg fusion is the prelude to the initial Ca2+ increase at fertilization in the mouse. , 1997, Development.

[25]  Kazuhiro Takahashi,et al.  Lowering intracellular and extracellular calcium contents prevents cytotoxic effects of ethylene glycol‐based vitrification solution in unfertilized mouse oocytes , 2004, Molecular reproduction and development.

[26]  D. Gardner,et al.  Addition of ascorbate during cryopreservation stimulates subsequent embryo development. , 2002, Human reproduction.

[27]  D. Gardner,et al.  A Laboratory Guide to the Mammalian Embryo , 2004 .

[28]  H. Wu,et al.  Injection of Sperm Cytosolic Factor Into Mouse Metaphase II Oocytes Induces Different Developmental Fates According to the Frequency of [Ca2+]i Oscillations and Oocyte Age1 , 2000, Biology of reproduction.

[29]  W. Son,et al.  High Survival Rates and Pregnancies of Human Oocytes Following Vitrification: Preliminary Report , 2005 .

[30]  J. Renard,et al.  Human embryo features that influence the success of cryopreservation with the use of 1,2 propanediol. , 1985, Fertility and sterility.

[31]  Sten Orrenius,et al.  Calcium: Regulation of cell death: the calcium–apoptosis link , 2003, Nature Reviews Molecular Cell Biology.

[32]  R. Schultz,et al.  Egg-to-embryo transition is driven by differential responses to Ca(2+) oscillation number. , 2002, Developmental biology.

[33]  M. Tucker,et al.  Effect of carrier system on the yield of human oocytes and embryos as assessed by survival and developmental potential after vitrification. , 2002, Reproduction.

[34]  Timothy D. Veenstra,et al.  Proteomic patterns: their potential for disease diagnosis , 2005, Molecular and Cellular Endocrinology.

[35]  N. Songsasen,et al.  Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. , 1996, Biology of reproduction.

[36]  S. Willadsen,et al.  Detrimental effects of sodium during mouse oocyte cryopreservation. , 1998, Biology of reproduction.

[37]  M. Nagano,et al.  Vitrification of mouse oocytes in ethylene glycol-raffinose solution: effects of preexposure to ethylene glycol or raffinose on oocyte viability. , 2001, Cryobiology.

[38]  T. Cheek,et al.  Cell cycle progression of parthenogenetically activated mouse oocytes to interphase is dependent on the level of internal calcium. , 1992, Journal of cell science.

[39]  G. Vajta,et al.  Highly efficient vitrification method for cryopreservation of human oocytes. , 2005, Reproductive biomedicine online.

[40]  J T Kline,et al.  Repetitive calcium transients and the role of calcium in exocytosis and cell cycle activation in the mouse egg. , 1992, Developmental biology.

[41]  K. Cha,et al.  Improved human oocyte development after vitrification: a comparison of thawing methods. , 1999, Fertility and sterility.

[42]  S. Emiliani,et al.  Comparison of ethylene glycol, 1,2-propanediol and glycerol for cryopreservation of slow-cooled mouse zygotes, 4-cell embryos and blastocysts. , 2000, Human reproduction.

[43]  T. Buschmann,et al.  Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI TOF-MS) and ProteinChip technology in proteomics research. , 2004, Pathology, research and practice.

[44]  C. Matthews,et al.  Freeze-thaw-induced changes of the zona pellucida explains decreased rates of fertilization in frozen-thawed mouse oocytes. , 1990, Journal of reproduction and fertility.

[45]  J. Yovich,et al.  Cryopreservation of oocytes and embryos: use of a mouse model to investigate effects upon zona hardness and formulate treatment strategies in an in-vitro fertilization programme. , 1997, Human reproduction.

[46]  C. Flamigni,et al.  Pregnancies and births after oocyte cryopreservation. , 2004, Fertility and sterility.

[47]  A. Trounson,et al.  Parthenogenetic activation of unfertilized mouse oocytes by exposure to 1,2-propanediol is influenced by temperature, oocyte age, and cumulus removal. , 1989, Gamete research.

[48]  D. Gardner,et al.  A proteomic analysis of mammalian preimplantation embryonic development. , 2005, Reproduction.

[49]  G. Vajta,et al.  Open pulled straw (OPS) vitrification: A new way to reduce cryoinjuries of bovine ova and embryos , 1998, Molecular reproduction and development.

[50]  K. P. Katayama,et al.  High survival rate of vitrified human oocytes results in clinical pregnancy. , 2003, Fertility and sterility.

[51]  J. Van der Elst,et al.  Parthenogenetic activation pattern and microtubular organization of the mouse oocyte after exposure to 1,2-propanediol. , 1992, Cryobiology.

[52]  L. Kuleshova,et al.  Vitrification can be more favorable than slow cooling. , 2002, Fertility and sterility.

[53]  R. E. Pitt,et al.  Cryopreservation of Drosophila melanogaster embryos , 1989, Nature.

[54]  S. Nakamura,et al.  Successful birth after transfer of vitrified human blastocysts with use of a cryoloop containerless technique. , 2001, Fertility and sterility.

[55]  T. Falcone,et al.  Gonadotropin-Releasing Hormone Agonists ( GnRH-a ) , 2004 .