Effect of N-acetyl cysteine on the quality of blastocyst formation rate using cultured vitrified murine embryos

PDF
DOI: https://doi.org/10.12681/jhvms.25078
Keywords:
N-acetylcysteine vitrification early embryos mouse blastocyst quality
S. SIGÜENZA
Department of Cellular Biology, School of Life Sciences, University of Extremadura, Badajoz, Spain
I.S. ÁLVAREZ
Department of Cellular Biology, School of Life Sciences, University of Extremadura, Badajoz, Spain
E. MATILLA
Department of Cellular Biology, School of Life Sciences, University of Extremadura, Badajoz, Spain
Abstract

Vitrification is the best method for embryo cryopreservation although it increases endogenous reactive oxygen species (ROS) production. N-acetylcysteine (NAC) a free radical scavenger may be used for reducing ROS toxic effects. The aim of the present study is to investigate potential beneficial effects of NAC on the developmental embryo competence applying different culture conditions in vitrified-warmed 2-cell embryos derived in vivo or in vitro. Thus, 2-cell embryos were vitrified or cultured fresh in presence or absence of 1 mM of NAC during: a) the entire embryo culture, b) for 24 hours with NAC at days 1.5 (G1) or 2.5 (G2) and returned to basal embryo culture (KSOM) or c) cultured in the presence of NAC for 12 hours at day 3.5 (G3). Despite NAC addition to fresh or vitrified embryos produced in vivo or by IVF, blastocyst rates remained unchanged. In vitrified-warmed IU or IVF-derived embryos, total cell number varied when NAC was added at day 1.5 although differences were not significant (60.1 ± 1.9 vs. 59.4 ± 1.3 for IU G1 and control respectively; and 59.3 ± 1.6 and 52.6 ± 3.0 IVF G1 and control respectively; mean cell number ± SEM, p > 0.05). It seems that the embryo culture medium supplementation with 1 mM of NAC in the first day after vitrification of development improves blastocyst quality of murine embryos and does not exert any beneficial effect at oyher culture points.

Article Details
  • Section
  • Research Articles
Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Authors who publish with this journal agree to the following terms:

· Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution Non-Commercial License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.

· Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g. post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.

· Authors are permitted and encouraged to post their work online (preferably in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.

Downloads
Download data is not yet available.
References
Ali, A.A., J.F. Bilodeau and M.A. Sirard (2002). “Antioxidant requirements for bovine oocytes varies during in vitro maturation, fertilization and development.” Theriogenology 59(3): 939-949.
Azadbakht, M. and M.R. Valojerdi (2008). “Development of vitrified-warmed mouse embryos co-cultured with polarized or non-polarized uterine epithelial cells using sequential culture media.” Journal of Assisted Reproduction and Genetics 25(6): 251-261.
Bedaiwy, M.A., T. Falcone, M.S. Mohamed, A.A.N. Aleem, R.K. Sharma, S.E. Worley, J. Thornton and A. Agarwal (2004). “Differential growth of human embryos in vitro: Role of reactive oxygen species.” Fertility and Sterility 82(3): 593-600.
Castillo-Martín, M., S. Bonet, R. Morató and M. Yeste (2014). “Comparative effects of adding β-mercaptoethanol or l-ascorbic acid to culture or vitrification–warming media on IVF porcine embryos.” Reproduction, Fertility and Development 26(6):875-882.
Covarrubias, L., D. Hernández-García, D. Schnabel, E. Salas-Vidal and S. Castro-Obregón (2008). “Function of reactive oxygen species during animal development: Passive or active?” Developmental Biology 320(1): 1-11.
ESHRE (2011). The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting†. Human Reproduction. Gao, C., H.-B. Han, X.-Z. Tian, D.-X. Tan, L. Wang, G.-B. Zhou, S.-E.
Zhu and G.-S. Liu (2012). “Melatonin promotes embryonic development and reduces reactive oxygen species in vitrified mouse 2-cell embryos.” J Pineal Res 52(3): 305-311.
Ghandy, N. and A.A. Karimpur Malekshah (2017). “Which Stage of Mouse Embryos Is More Appropriate for Vitrification?” International Journal of Fertility & Sterility 10(4): 357-362.
Ghandy, N., Karimpur, M., Abbas, A. (2017). “Which Stage of Mouse Embryos Is More Appropriate for Vitrification?” International Journal of Fertility & Sterility 10(4): 357-362.
Guerin, P., S. El Mouatassim and Y. Menezo (2001). “Oxidative stress and protection against reactive oxygen species in the pre-implantation embryo and its surroundings.” Hum Reprod Update 7.
Guérin, P., S. El Mouatassim and Y. Ménézo (2001). “Oxidative stress and protection against reactive oxygen species in the pre-implantation embryo and its surroundings.” Hum Reprod Update 7(2): 175-189.
Jurisicova, A., I. Rogers, A. Fasciani, R.F. Casper and S. Varmuza (1998). “Effect of maternal age and conditions of fertilization on programmed cell death during murine preimplantation embryo development.” Mol Hum Rep 2: 139-145.
Kong, X., S. Yang, F. Gong, C. Lu, S. Zhang, G. Lu and G. Lin (2016). “The Relationship between Cell Number, Division Behavior and Developmental Potential of Cleavage Stage Human Embryos: A Time-Lapse Study.” PLoS One 11(4): e0153697.
Kuleshova, L.L. and A. Lopata (2002). “Vitrification can be more favorable than slow cooling.” Fertility and Sterility 78(3): 449-454.
Liang, Y., F.Y. Ning, W.J. Du, C.S. Wang, S.H. Piao and T.Z. An (2012). “The type and extent of injuries in vitrified mouse oocytes.” Cryobiology 64(2): 97-102.
Mallol, A., J. Santaló and E. Ibáñez (2013). “Comparison of three differential mouse blastocyst staining methods.” Systems Biology in Reproductive Medicine 59(2): 117-122.
Mandawala, A.A., S.C. Harvey, T.K. Roy and K.E. Fowler (2016). “Cryopreservation of animal oocytes and embryos: Current progress and future prospects.” Theriogenology 86(7): 1637-1644.
Marí, M., A. Morales, A. Colell, C. García-Ruiz and J.C. Fernández-Checa (2009). “Mitochondrial Glutathione, a Key Survival Antioxidant.” Antioxidants & Redox Signaling 11(11): 2685-2700.
Matilla, E., F.E. Martin-Cano, L. González-Fernandez, F.M. Sanchez-Margallo, I.S. Alvarez and B. Macias-Garcia (2019). “N-acetylcysteine addition after vitrification improves oocyte mitochondrial polarization status and the quality of embryos derived from vitrified murine oocytes.” BMC Veterinary Research 15(1): 31.
Momozawa, K., A. Matsuzawa, Y. Tokunaga, S. Abe, Y. Koyanagi, M. Kurita, M. Nakano and T. Miyake (2017). “Efficient vitrification of mouse embryos using the Kitasato Vitrification System as a novel vitrification device.” Reproductive Biology and Endocrinology 15(1):29.
Pedro, P.B., E. Yokoyama, S.E. Zhu, N. Yoshida, D.M. Valdez Jr, M. Tanaka, K. Edashige and M. Kasai (2005). “Permeability of Mouse Oocytes and Embryos at Various Developmental Stages to Five Cryoprotectants.” Journal of Reproduction and Development 51(2): 235- 246.
Rall W.F., F.G.M. (1985). “Ice-free cryopreservation of mouse embryos at -196 degrees C by vitrification.” Nature 313(6003): 3.
Rall, W.F. (1987). “Factors affecting the survival of mouse embryos cryopreserved by vitrification.” Cryobiology 24(5): 387-402.
Sawicki, W. and E.T. Mystkowska (1990). “In vivo staining of mouse preimplantation embryos with hoechst 33342.” The Anatomical Record 227(3): 359-362.
Silva, E., A.F. Greene, K. Strauss, J.R. Herrick, W.B. Schoolcraft and R.L. Krisher (2015). “Antioxidant supplementation during in vitro culture improves mitochondrial function and development of embryos from aged female mice.” Reproduction, Fertility and Development 27(6):975-983.
Somfai, T., M. Ozawa, J. Noguchi, H. Kaneko, N.W. Kuriani Karja, M. Farhudin, A. Dinnyés, T. Nagai and K. Kikuchi (2007). “Developmental competence of in vitro-fertilized porcine oocytes after in vitro maturation and solid surface vitrification: Effect of cryopreservation on oocyte antioxidative system and cell cycle stage.” Cryobiology 55(2): 115-126.
Somoskoi, B., N.A. Martino, R.A. Cardone, G.M. Lacalandra, M.E. Dell’Aquila and S. Cseh (2015). “Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification.” Reprod Biol Endocrinol 13: 22.
Tatone, C., G. Di Emidio, M. Vento, R. Ciriminna and P.G. Artini (2010). “Cryopreservation and oxidative stress in reproductive cells.” Gynecological Endocrinology 26(8): 563-567.
Tsang W.H. and Chow L.K. (2009). “Mouse embryo cryopreservation utilizing a novel high-capacity vitrification spatula.” Biotechniques 46(7): 3.
Vajta, G. (2000). “Vitrification of the oocytes and embryos of domestic animals.” Animal Reproduction Science 60-61: 357-364.
Van der Elst J, Amerijckx Y and V.S. A. (1998). “Ultra-rapid freezing of mouse oocytes lowers the cell number in the inner cell mass of 5 day old in-vitro cultured blastocysts.” Human Reproduction 13(6): 5.
Yan, C.L., X.W. Fu, G.B. Zhou, X.M. Zhao, L. Suo and S.E. Zhu (2010). “Mitochondrial behaviors in the vitrified mouse oocyte and its parthenogenetic embryo: effect of Taxol pretreatment and relationship to competence.” Fertil Steril 93(3): 959-66.
Zhang, J., J. Cui, X. Ling, X. Li, Y. Peng, X. Guo, B.C. Heng and G.Q. Tong (2009). “Vitrification of mouse embryos at 2-cell, 4-cell and 8-cell stages by cryotop method.” Journal of Assisted Reproduction and Genetics 26(11-12): 621-628.