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Detrimental effects of antibiotics on mouse embryos in chromatin integrity, apoptosis and expression of zygotically activated genes*

  • Jun Liu (a1), Shuang Tang (a1), Wei Xu (a1), Yongsheng Wang (a1), Baoying Yin (a1) and Yong Zhang (a2) (a1)...

The effects of specific components in culture medium on embryo physiology have been extensively investigated to optimize in vitro culture systems; however, little attention has been paid to antibiotics, the reagents used most commonly in culture systems to prevent contamination. To investigate the potential effects of routine use of antibiotics on cultured embryos, mouse zygotes were cultured with or without antibiotics. In both groups, the developmental rate and cell number of blastocysts appear to be normal. The proportion of embryos with blastomere fragmentation increased slightly when embryos were cultured with antibiotics. In contrast, the presence of antibiotics in the embryo culture system significantly disturbs expression of zygotically activated genes, damages chromatin integrity and increases apoptosis of cultured embryos. These results provide evidence that, when cultured with antibiotics, embryos with normal appearance may possess intrinsic physiological and genetic abnormalities. We demonstrate that the adverse effects of antibiotics on mammalian embryos are more severe than we previously presumed and that antibiotics are not essential for sterility of embryo culture system therefore abolishing antibiotic supplementation during embryo culture.

Corresponding author
All correspondence to: Yong Zhang. Institute of Biotechnology, Northwest A&F University, Yangling, Shaanxi 712100, P. R. China. Fax: +86 02987080085. e-mail:
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This work is supported by grants from the Important National Science & Technology Specific Projects, China (No. 2008ZX08007-004).

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Aitken R.J. (2008). Just how safe is assisted reproductive technology for treating male factor infertility? Expert Rev. Obstet. Gynecol. 3, 267–71.
Amonn F., Baumann U., Wiesmann U.N., Hofmann K. & Herschkowitz N. (1978). Effects of antibiotics on the growth and differentiation in dissociated brain cell cultures. Neuroscience 3, 465–8.
Bavister B.D. (1995). Culture of preimplantation embryos: facts and artifacts. Hum. Reprod. Update 1, 91148.
Bavister B.D. (2000). Interactions between embryos and the culture milieu. Theriogenology 53, 619–26.
Christians E., Campion E., Thompson E.M. & Renard J-P. (1995). Expression of the HSP 70.1 gene, a landmark of early zygotic activity in the mouse embryo, is restricted to the first burst of transcription. Development 121, 113–22.
Christians E., Michelob E. & Renard J-P. (1997). Hsp70 genes and heat shock factors during preimplantation phase of mouse development. Cell. Mol. Life Sci. 53, 168–78.
Ericson A. & Kallen B. (2001). Congenital malformations in infants born after IVF: a population-based study. Hum. Reprod. 16, 504–9.
Felsenfeld G., Boyes J., Chung J., Clark D. & Studitsky V. (1996). Chromatin structure and gene expression. Proc. Natl. Acad. Sci. USA 93, 9384–8.
Fernandez-Gonzalez R., Moreira P., Bilbao A., Jimenez A., Perez-Crespo M., Ramirez M.A., De Fonseca F.R., Pintado B. & Gutierrez-Adan A. (2004). Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior. Proc. Natl. Acad. Sci. USA 101, 5880–5.
Fiorenza M.T., Bevilacqua A., Canterini S., Torcia S., Pontecorvi M. & Mangia F. (2004). Early transcriptional activation of the Hsp70.1 gene by osmotic stress in one-cell embryos of the mouse. Biol. Reprod. 70, 1606–113.
Gandolfi F. (1994). Autocrine, paracrine and environmental-factors influencing embryonic-development from zygote to blastocyst. Theriogenology 41, 95100.
Gjorret J.O., Knijn H.M., Dieleman S.J., Avery B., Larsson L.I. & Maddox-Hyttel P. (2003). Chronology of apoptosis in bovine embryos produced in vivo and in vitro. Biol. Reprod. 69, 1193–200.
Hansen M., Kurinczuk J.J., Bower C. & Webb S. (2002). The risk of major birth defects after intracytoplasmic sperm injection and in vitro fertilization. N. Engl. J. Med. 346, 725–30.
Hansen M., Bower C., Milne E., de Klerk N. & Kurinczuk J.J. (2005). Assisted reproductive technologies and the risk of birth defects – a systematic review. Hum. Reprod. 20, 328–38.
Hardy K. (1997). Cell death in the mammalian blastocyst. Mol. Hum. Reprod. 3, 919–25.
Ho Y., Wigglesworth K., Eppig J.J. & Schultz R.M. (1995). Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol. Reprod. Dev. 41, 232–8.
Jurisicova A., Varmuza S. & Casper R.F. (1996). Programmed cell death and human embryo fragmentation. Mol. Hum. Reprod. 2, 93–8.
Kaneko K.J., Cullinan E.B., Latham K.E. & DePamphilis M.L. (1997). Transcription factor mTEAD-2 is selectively expressed at the beginning of zygotic gene expression in the mouse. Development 124, 1963–73.
Kanka J. (2003). Gene expression and chromatin structure in the preimplantation embryo. Theriogenology 59, 319.
Khosla S., Dean W., Brown D., Reik W. & Feil R. (2001a). Culture of preimplantation mouse embryos affects fetal development and the expression of imprinted genes. Biol. Reprod. 64, 918–26.
Khosla S., Dean W., Reik W. & Feil R. (2001b). Culture of preimplantation embryos and its long-term effects on gene expression and phenotype. Hum. Reprod. Update 7, 419–27.
Kigami D., Minami N., Takayama H. & Imai H. (2003). MuERV-L is one of the earliest transcribed genes in mouse one-cell embryos. Biol. Reprod. 68, 651–4.
Knijn H.M., Gjorret J.O., Vos P.L., Hendriksen P.J., Van Der Weijden B.C., Maddox-Hyttel P. & Dieleman S.J. (2003). Consequences of in vivo development and subsequent culture on apoptosis, cell number, and blastocyst formation in bovine embryos. Biol. Reprod. 69, 1371–8.
Lane M. & Gardner D.K. (2003). Ammonium induces aberrant blastocyst differentiation, metabolism, pH regulation, gene expression and subsequently alters fetal development in the mouse. Biol. Reprod. 69, 1109–17.
Livak K.J., Flood S.J.A., Marmaro J., Giusti W. & Deetz K. (1995). Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic-acid hybridization. Genome Res. 4, 357–62.
Magli M. C., Gianaroli L., Fiorentino A., Ferraretti A. P., Fortini D. & Panzella S. (1996). Improved cleavage rate of human embryos cultured in antibiotic-free medium. Hum. Reprod. 11, 1520–4.
Mann M.R., Lee S.S., Doherty A.S., Verona R.I., Nolen L.D., Schultz R.M. & Bartolomei M.S. (2004). Selective loss of imprinting in the placenta following preimplantation development in culture. Development 131, 3727–35.
Manuvakhova M., Keeling K. & Bedwell D.M. (2000). Aminoglycoside antibiotics mediate context-dependent suppression of termination codons in a mammalian translation system. RNA 6, 1044–55.
McKieman S.H. & Bavister B.D. (1990). Environmental variables influencing in vitro development of hamster 2-cell embryos to the blastocyst stage. Biol. Reprod. 43, 404–13.
McKieman S.H. & Bavister B.D. (2000). Culture of one-cell hamster embryos with water soluble vitamins: pantothenate stimulates blastocyst production. Hum. Reprod. 15, 157–64.
Niemann H. & Wrenzycki C. (2000). Alterations of expression of developmentally important genes in preimplantation bovine embryos by in vitro culture conditions: implications for subsequent development. Theriogenology 53, 2134.
Olson C.K., Keppler-Noreuil K.M., Romitti P.A., Budelier W.T., Ryan G., Sparks A.E. & Van Voorhis B.J. (2005). In vitro fertilization is associated with an increase in major birth defects. Fertil. Steril. 84, 1308–15.
Paria B.C. & Dey S.K. (1990). Preimplantation embryo development in vitro: cooperative interactions among embryos and role of growth factors. Proc. Natl. Acad. Sci. USA 87, 4756–60.
Rizos D., Gutierrez-Adan A., Perez-Garnelo S., De La Fuente J., Boland M.P. & Lonergan P. (2003). Bovine embryo culture in the presence or absence of serum: implications for blastocyst development, cryotolerance, and messenger RNA expression. Biol. Reprod. 68, 236–43.
Schultz R.M. (1993). Regulation of zygotic gene activation in the mouse. Bioessays 15, 531–8.
Schultz R.M. & Williams C.J. (2002). The science of ART. Science 296, 2188–90.
Schultz R.M. & Worrad D.M. (1995). Role of chromatin structure in zygotic gene activation in the mammalian embryo. Semin. Cell Biol. 6, 201–8.
Schultz R.M., Davis W., Stein P. & Svoboda P. (1999). Reprogramming of gene expression during preimplantation development. J. Exp. Zool. 285, 276–82.
Stemp G., Pascoe S. & Gatehouse D. (1989). In vitro and in vivo cytogenetic studies of three beta-lactam antibiotics (penicillin VK, ampicillin and carbenicillin). Mutagenesis 4, 439–45.
Sturmey R.G., Brison D.R. & Leese H.J. (2008). Assessing embryo viability by measurement of amino acid turnover. Reprod. Biomed. Online 17, 486–96.
Sturmey R.G., Hawkhead J.A., Barker E.A. & Leese H.J. (2009). DNA damage and metabolic activity in the preimplantation embryo. Hum. Reprod. 24, 8191.
Summers M. & Biggers J.D. (2003). Chemically defined media and the culture of mammalian preimplantation embryos: historical perspective and current issues. Hum. Reprod. Update 9, 557–82.
Suzuki T., Minami N., Kono T. & Imai H. (2006). Zygotically activated genes are suppressed in mouse nuclear transferred embryos. Cloning Stem Cells 8, 295304.
Takenaka M., Horiuchi T. & Yanagimachi R. (2007). Effects of light on development of mammalian zygotes. Proc. Natl. Acad. Sci. USA 104, 14289–93.
Tang S., Wang Y., Zhang D., Gao Y., Ma Y., Yin B., Sun J., Liu J. & Zhang Y. (2009). Reprogramming donor cells with oocyte extracts improves in vitro development of nuclear transfer embryos. Anim. Reprod. Sci. 115, 19.
Vergouw C.G., Botros L.L., Roos P., Lens J.W., Schats R., Hompes P.G., Burns D.H. & Lambalk C.B. (2008). Metabolomic profiling by near-infrared spectroscopy as a tool to assess embryo viability: a novel, non-invasive method for embryo selection. Hum. Reprod. 23, 1499–504.
Vinson R.K. & Hales B.F. (2002). DNA repair during organogenesis. Mutat. Res. 509, 7991.
Watkins A.J., Platt D., Papenbrock T., Wilkins A., Eckert J.J., Kwong W.Y., Osmond C., Hanson M. & Fleming T.P. (2007). Mouse embryo culture induces changes in postnatal phenotype including raised systolic blood pressure. Proc. Natl. Acad. Sci. USA 104, 5449–54.
Xie Y., Wang F., Zhong W., Puscheck E., Shen H. & Rappolee D.A. (2006). Shear stress induces preimplantation embryo death that is delayed by the zona pellucida and associated with stress-activated protein kinase-mediated apoptosis. Biol. Reprod. 75, 4555.
Zhou H., McKiernan S.H., Ji W. & Bavister B.D. (2000). Effect of antibiotics on development in vitro of hamster pronucleate ova. Theriogenology 54, 9991006.
Zhu Y., Carroll M., Papa F.R., Hochstrasser M. & D'Andrea A.D. (1996). DUB-1, a deubiquitinating enzyme with growth-suppressing activity. Proc. Natl. Acad. Sci. USA 93, 3275–9.
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