Purpose
A pilot study was performed to test the diagnostic value of in vitro DNA fluorescence in oocytes that failed to fertilize after IVF. Ten patients with a cleavage rate less than 20% after IVF were included.
Results
Uncleaved oocytes were observed by fluorescence microscopy after incubation with the DNA fluorescent dye Hoechst 33342. Four main causes which may have contributed to the low cleavage rate were found: (1) sperm incapacity to penetrate the oocyte despite the absence of the usual criteria for male infertility, (2) oocyte immaturity, (3) delayed fertilization, and (4) oocyte abnormalities revealed by aberrations in the morphology of the female chromatin.
Conclusions
The possibility of a rapid and detailed analysis of the maturational status of unfertilized oocytes, the morphology of the female chromatin, the presence and quantity of spermatozoa tightly bound to the zona pellucida, and sperm penetration into the oocyte without subsequent pronucleus formation, using DNA fluorescence, allows us to clarify further the cause of fertilization failure and to orient infertility treatment toward the male, the female, or both partners.
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References
Bomsel-Helmreich O, Durand-Gasselin I, Pennehouat G, Vu N Huyen L, Antoine JM, Salat-Baroux J: Structure and ultrastructure of human preovulatory oocytes immediately before the administration of hCG.In From Ovulation to Implantation, JLH Evers, MJ Heineman (eds). Exerpta Medica, Int. Congr. Ser. 917, 1990, pp 81–90
Sathananthan AH: Maturation of the human oocyte in vitro: Nuclear events during meiosis (an ultrastructural study). Gamete Res 1985;12:237–254
Van Blerkom J, Henry G: Cytogenetic analysis of living human oocytes: Cellular basis and developmental consequences of pertubations in chromosomal organisation and complement. Hum Reprod 1988;3:777–790
Wissen AMB VAN, Bomsel-Helmreich O, Debey P, Eisenberg C, Vaulthier, D, Pennehouat G: Fertilization and ageing processes in non divided human oocytes after GnRHa treatment: An analysis of individual oocytes. Hum Reprod 1991;6:879–884
Tarkowski AK: An air-drying method for chromsome preparation from mouse eggs. Cytogenetics 1966;5:394–400
Schmiady H, Kentenich H: Premature chromosome condensation after in vitro fertilization. Hum Reprod 1989;4:689–695
Pieters MHEC, Geraedts JPM, Dumoulin JCM, Evers JLH, BRAS M, Kornips FHAC, Menheere PPCA: Cytogenetic analysis of in vitro fertilization (IVF) failures. Hum Genet 1989;81:367–370
Selva J, Martin-Pont B, Hugues JN, Rince P, Fillion C, Herve F, Tamboise A, Tamboise E: Cytogenetic study of human oocytes uncleaved after in-vitro fertilization. Hum Reprod 1991;6:709–713
Tarin JJ, Ruiz A, Miro F, Bonilla-Musoles F, Pellicer A: Failed in vitro fertilization of human oocytes: a cytogenetic analysis. Fertil Steril 1991;56:290–295
World Health Organization: Laboratory Manual for the Examination of Human Semen and Semen-Cervical Mucus Interaction, MA Belsey, R Eliasson, AJ Gallegos, KS Moghissi, CA Paulson (eds). Singapore, Press Concern, 1980
Frydman R, Parneix I, Belaisch-Allart J, Forman R, Hazout A, Fernandez H, Testart J: LHRH agonists in IVF: Different methods of utilization and comparison with previous ovulation stimulation treatments. Hum Reprod 1988;3:559–561
Debey P, Renard JP, Coppey-Moisan M, Monnot I, Geze M: Dynamics of chromatin changes in live one-cell mouse embryos: a continuous follow-up by fluorescence microscopy. Exp Cell Res 1989;183:413–433
Gwatkin RBL, Conover JC, Collins RL, Quigley MM: Failed fertilization in human in vitro fertilization analysed with the deoxyribonucleic acid-specific fluorochrome Hoechst 33342. Am J Obstet Gynecol 1989;160:31–35
Liu DY, Lopata A, Johnston WIH, Baker HWG: Human sperm-zona pellucida binding, sperm characteristics and in vitro fertilization. Hum Reprod 1989;4:696–701
Ron-El R, Nachum H, Herman A, Golan A, Caspi E, Soffer Y: Delayed fertilization and poor embryonic development associated with impaired semen quality. Fertil Steril 1991;55:338–344
Liu DY, Lopata A, Johnston WIH, Baker HWG: A human sperm-zona pellucida binding test using oocytes that failed to fertilize in vitro. Fertil Steril 1988;50:782–788
Van Wissen B, Bomsel-Helmreich O, de Ziegler D, Pietri V, Frydman R: Examen morpho-fonctionnel des gamètes en cas d'échec de fécondation in vitro. Numéro spécial de Fertilité Contraception Sexualité, 1:1992;20:801–803
Mahadevan MM, Trounson AO, Wood C, Leeton JF: Effect of oocyte quality and sperm characteristics on the number of spermatozoa bound to the zona pellucida of human oocytes inseminated in vitro. J Vitro Fert Embryo Transfer 1987;4:223–227
Kruger TF, Acosta AA, Simmons KF, Swanson RJ, Matta JF, Oehninger S: Predictive value of abnormal sperm morphology in vitro fertilization. Fertil Steril 1988;49:112–117
Liu DY, Du Plessis Y, Nayudu PL, Johnston WIH, Baker HWG: The use of in vitro fertilization to evaluate putative tests of human sperm function. Fertil Steril 1988;49:272–279
Burkman LJ, Coddington CC, Franken DR, Kruger T, Rosenwaks Z, Hodgen GD: The hemi-zona assay (HZA): Development of a diagnostic test for the binding of human spermatozoa to the human hemizona pellucida to predict fertilization potential. Fertil Steril 1988;49:688–697
Kruger TF, Swanson RJ, Hamilton M, Simmons KF, Acosta AA, Matta JF, Oehninger S, Morsedhi M: Abnormal sperm morphology and other semen parameters related to the outcome of the hamster oocyte human sperm penetration assay. Int J Androl 1988;11:107–113
Lopata A, Leung PC: The fertilizability of human oocytes at different stages of meiotic maturation.In IVF and Other Assisted Reproduction. HW Jones, CS Schroeder (eds). Ann NY Acad Sci, 1988, pp 324–336
Kubiak J: Mouse oocytes gradually develop the capacity for activation during the metaphase II arrest. Dev Biol 1989;136:537–545
Zenzes MT, Belkien L, Bordt J, Kan I, Schneider HPG, Nieschlag E: Cytologic investigation of human in vitro fertilization failures. Fertil Steril 1985;43:883–891
Jamieson ME, Fleming R, Kader S, Ross KS, Yates RWS, Coutts JRT: In vivo and in vitro maturation of human oocytes: Effects on embryo development and polyspermic fertilization. Fertil Steril 1991;56:93–97
Trounson AO, Mohr LR, Wood C, Leeton JF: Effect of delayed insemination on in-vitro fertilization, culture and transfer of human embryos. J Reprod Fert 1982;64:285–294
Trounson A, Webb J: Fertilization of human oocytes following reinsemination in vitro. Fertil Steril 1984;41:816–819
Pampiglione JS, Mills C, Campbell S, Steer C, Kingsland C, Mason B: The clinical outcome of reinsemination of human oocytes fertilized in vitro. Fertil Steril 1990;53:306–310
Oehninger S, Acosta AA, Veeck LL, Simonetti S, Muasher SJ: Delayed fertilization during in vitro fertilization and embryo transfer cycles: Analysis of causes and impact on overall results. Fertil Steril 1989;52:991–997
Veeck LL, Wortham JWE, Witmeyer J, Sandow BA, Acosta AA, Garcia JE, Jones GS, Jones HW: Maturation and fertilization of morphologically immature human oocytes in a program of in vitro fertilization. Fertil Steril 1983;39:594–602
Rudak E, Dor J, Kimchi M, Goldman B, Levran D, Mashiach S: Anomalies of human oocytes from infertile women undergoing treatment by in vitro fertilization. Fertil Steril 1990;54:292–296
Edwards RG: Maturation in vitro of human ovarian oocytes. Lancet 1965;2:926–929
Nayudu PL, Lopata A, Jones GM, Gook DA, Bourne HM, Sheather SJ, Brown TC, Johnston WIH: An analysis of human oocytes and follicles from stimulated cycles: Oocyte morphology and associated follicular fluid characteristics. Hum Reprod 1989;4:558–567
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Van Wissen, B., Eisenberg, C., Debey, P. et al. In vitro DNA fluorescence after in vitro fertilization (IVF) failure. J Assist Reprod Genet 9, 564–571 (1992). https://doi.org/10.1007/BF01204255
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DOI: https://doi.org/10.1007/BF01204255