Skip to main content
SpringerLink
Log in

CLINICAL ASSISTED REPRODUCTION: Updating Quality Control Assays in the Assisted Reproductive Technologies Laboratory with a Cryopreserved Hamster Oocyte DNA Cytogenotoxic Assay

  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose: Despite advances in assisted reproduction, there is no progress in quality control bioassays. The objectives were to develop a comet assay to measure DNA fragmentation in thawed cryopreserved oocytes and compare this assay with one-cell mouse embryo bioassay.

Methods: Thawed hamster oocytes from a commercial source were incubated in culture media with either 0-, 50-, or 100-μM hydrogen peroxide, or, in media exposed to different contact materials and unknown proficiency analytes. Incubation time was 1.5 h at 37°C. The oocytes were dried, fixed, stained with acridine orange, embedded in a mini-agarose layer and electrophoresis was carried out. Fluorescent images were analyzed. The results were compared with standard one-cell mouse assay data.

Results: The 100-μM hydrogen peroxide treatment caused greatest DNA fragmentation in the hamster oocytes at Hours 1 and 2. A dose response was observed. Intrassay coefficient of variation was 5.7%. Only one of the five materials tested passed both assays. The data for the unknown proficiency analytes were similar for both assays.

Conclusions: The oocyte comet assay demonstrated DNA fragmentation in the presence of toxic substances. The detection of toxicity in two materials that passed the mouse bioassay suggested increased sensitivity in the new assay. The oocyte comet assay and the mouse bioassay results matched in the proficiency test. However, more studies are still needed to determine optimal sensitivity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. Ackerman SB, Swanson RJ, Stokes GK, Veeck LL: Culture of mouse preimplantation embryos as a quality control assay for human in vitro fertilization. Gamete Res 1984;9:145-152

    Google Scholar 

  2. Condon-Mahoney M, Wortham JWE Jr, Bundren JC, Witmyer J, Shirley B: Evaluation of human fetal cord sera, Ham's F-10 medium, and in vitro culture materials with a mouse in vivo fertilization system. Fertil Steril 1985;44:521-525

    Google Scholar 

  3. Fukuda A, Noda Y, Tsukui S, Matsumoto H, Yano J, Mori T: Influences of water quality on in vitro fertilization and embryo development for the mouse. J Vitro Fert Embryo Transf 1987;4:40-45

    Google Scholar 

  4. Parinaud J, Reme J, Monrozies X, Favrin S, Sarramon M, Pontonnier G: Mouse system quality control is necessary before the use of new material for in vitro fertilization and embryo transfer. J Vitro Fert Embryo Transf 1987;4:56-58

    Google Scholar 

  5. Davidson A, Vremiseh M, Lobo RA, Paulson RJ: Mouse embryo culture as quality control for human in vitro fertilization: the one-cell versus two-cell model. Fertil Steril 1988;49:516-521

    Google Scholar 

  6. Rinehart JS, Bavister BD, Gerrity M: Quality control in the in vitro fertilization laboratory: comparison of bioassay systems for water quality. J Vitro Fert Embryo Transf 1988;5:335-342

    Google Scholar 

  7. Gorrill MJ, Rinehart JS, Tamhane AC, Gerrity M: Comparison of the hamster sperm motility assay to the mouse one-cell and two-cell bioassays as quality control tests for in vitro fertilization. Fertil Steril 1991;55:345-354

    Google Scholar 

  8. Johnson DE, Hodgen GD:Syringe-associated toxicity of culture media on mouse and monkey preembryos. J Vitro Fert Embryo Transf 1991;8:198-201

    Google Scholar 

  9. Bavister BD, Andrews JC: A rapid sperm motility bioassay procedure for quality-control testing of water and culture media. J Vitro Fert Embryo Transf 1988;5:67-75

    Google Scholar 

  10. Stewart-Savage J, Bavister BD: Deterioration of stored culture media as monitored by a sperm motility bioassay. J Vitro Fert Embryo Transf 1988;5:76-80

    Google Scholar 

  11. Purdy JM: Methods for fertilization and embryo culture in vitro. In Human Conception In Vitro RG Edwards, JM Purdy (eds), London, Academic Press, 1982, pp. 135-148

    Google Scholar 

  12. Critchlow JD, Matson PL, Newman MC, Horne G, Troup SA, Lieberman BA: Quality control in an in-vitro fertilization laboratory: Use of human sperm survival studies. Hum Reprod 1989;4:545-549

    Google Scholar 

  13. Bertheussen K, Holst N, Fosdahl F, Høie KE: A new cell culture assay for quality control in IVF. Hum Reprod 1989;4:531-535

    Google Scholar 

  14. Fleming TP, Pratt HP, Braude PR: The use of mouse preimplantation embryos for quality control of culture reagents in human in vitro fertilization programs: A cautionary note. Fertil Steril 1987;47:858-860

    Google Scholar 

  15. Silverman IH, Cook CL, Sanfilippo JS, Yussman MA, Schultz GS, Hilton FK: Ham's F-10 constituted with tap water supports mouse conceptus development in vitro. J Vitro Fert Embryo Transf 1987;4:185-187

    Google Scholar 

  16. Östling O, Johanson KJ: Microelectrophoretic study of radiation-induced DNA damages in individual cells. Biochem Biophys Res Commun 1984;123:291-298

    Google Scholar 

  17. Singh NP, McCoy MT, Tice RR, Schneider EL: A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988;175:184-191

    Google Scholar 

  18. Singh NP, Danner DB, Tice RR, McCoy MT, Collins GD, Schneider EL: Abundant alkali sensitive sites in DNA of human and mouse sperm. Exp Cell Res 1989;184:461-470

    Google Scholar 

  19. Kent CR, Eady JJ, Ross GM, Steel GG: The comet moment as a measure of DNA damage in the comet assay. Int J Radiat Biol 1995;67:655-660

    Google Scholar 

  20. Klaude M, Ericksson S, Nygren J, Ahnström G: The comet assay: Mechanisms and technical considerations. Mutat Res 1996;363:89-96

    Google Scholar 

  21. Rafferty KA Jr.: Superovulation and phasing of ovulation. In Methods in Experimental Embryology of the Mouse, KA Rafferty Jr. (eds), Baltimore, The John Hopkins Press, 1970, pp. 23-29

    Google Scholar 

  22. Fairbairn DW, Olive PL, O'Neill KL: The comet assay: A comprehensive review. Mutat Res 1995;339:37-59

    Google Scholar 

  23. McKelvey-Martin VJ, Green MHL, Schmezer P, Pool-Zobel BL, De Meo MP, Collins A: The single cell gel electrophoresis assay (comet assay): A European review. Mutat Res 1993;288:47-63

    Google Scholar 

  24. Hughes CM, Lewis SE, McKelvey-Martin VJ, Thompson W: A comparison of baseline and induced DNA damage in human spermatozoa from fertile and infertile men, using a modified comet assay. Mol Hum Reprod 1996;2:613-619

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, California, 92350

    Philip J. Chan, J. Hazel Calinisan, Johannah U. Corselli, William C. Patton & Alan King

  2. Department of Physiology and Pharmacology, Loma Linda University School of Medicine, Loma Linda, California, 92350

    Philip J. Chan

Authors
  1. Philip J. Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Hazel Calinisan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Johannah U. Corselli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. William C. Patton
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alan King
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chan, P.J., Calinisan, J.H., Corselli, J.U. et al. CLINICAL ASSISTED REPRODUCTION: Updating Quality Control Assays in the Assisted Reproductive Technologies Laboratory with a Cryopreserved Hamster Oocyte DNA Cytogenotoxic Assay. J Assist Reprod Genet 18, 129–134 (2001). https://doi.org/10.1023/A:1009472323011

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009472323011

Search

Navigation