Evaluation of synthetic serum substitute versus serum as protein supplementation for mouse and human embryo culture

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Synthetic serum substitute (SSS) supported superior mouse embryo development and comparable human embryo development to serum, with a trend towards better fertilization rates in SSS.

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The study evaluated whether Synthetic Serum Substitute (SSS) can replace serum protein in embryo culture, comparing SSS with human serum in two experiments. In Experiment I, 1-cell mouse embryos were cultured with 15% protein supplementation from either no protein, serum from women with endometriosis or tubal infertility, fertile donor serum, or 15% SSS, and embryo development was assessed up to 96 hours; the authors reported no major caveat in the abstract regarding limitations. SSS supported substantially higher morula formation (64.9% at 48 hours) and blastocyst development (29.7% at 72 hours, 83.7% by 96 hours) than the serum conditions evaluated, and blastocyst hatching was observed in the SSS group (43.2% by 96 hours), with serum group hatching largely absent. In Experiment II with human IVF embryos, embryo stage/quality did not differ between SSS and serum, though fertilization rates tended to be higher with SSS (70.0% vs 55.0%, P=0.07). Relevance to endometriosis: the paper includes serum from women with endometriosis as one of the serum comparison groups in the mouse embryo experiments, finding that endometriosis serum performed worse than SSS for blastocyst development, though the paper’s main focus is comparing SSS versus serum for embryo culture.

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Abstract

PurposeOur purpose was to determine the effect of Synthetic Serum Substitute (SSS) versus serum supplementation on fertilization rates and subsequent development of embryos from patients undergoing IVF.ProcedureExperiment I compared the effects of SSS to human serum on mouse embryo development. Two hundred one-cell B6D2F1 mouse embryos were cultured in 100-microliter droplets of human tubal fluid (HTF) containing either (1) no protein (control; n = 37), (2) 15% serum from women with tubal infertility (n = 44), (3) 15% serum from women with endometriosis (n = 49), (4) 15% fertile donor serum (n = 33), or (5) 15% SSS (n = 37). Experiment II compared the effects of SSS to human serum on the development of embryos from patients undergoing IVF. Thirty-three women were included in this study. A total of 371 oocytes was cultured in HTF containing either (1) maternal or donor serum (n = 140) or (2) 15% SSS (n = 231). Embryo development was evaluated 48 hr after fertilization.ResultsIn Experiment I, the rate of blastocyst development was evaluated at 48, 72, and 96 hr of culture. Sixty-four and nine-tenths percent of embryos cultured in SSS were morulae at 48 hr of culture (versus 5.4, 0, 8.2, and 6.1 in Groups 1, 2, 3, and 4, respectively). By 72 hr, 29.7% of these embryos had developed into blastocysts (versus 0, 0, 8.2, and 3.0, for Groups 1, 2, 3, and 4, respectively). This percentage increased to a total of 83.7 after 96 hr (versus 27.0, 20.4, 38.8, and 39.4 for Groups 1, 2, 3, and 4, respectively). Forty-three and two-tenths percent of the blastocysts cultured in SSS had hatched from their zonae by 96 hr. With the exception of Group 5, which had a rate of 9.1%, embryo hatching was not observed in any of the groups at the termination of culture (96 hr). In Experiment II there were no differences in cell stage or quality of human embryos cultured in SSS or serum, but fertilization rates tended to be better (P = 0.07) for oocytes inseminated in media containing SSS (70.0%, vs 55.0% for serum).ConclusionsSSS appears to be a superior protein source for mouse embryo growth and is as good as serum from fertile donors in promoting in vitro human embryo development.
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Abstract

Purpose: Our purpose was to determine the effect of Synthetic Serum Substitute (SSS) versus serum supplementation on fertilization rates and subsequent development of embryos from patients undergoing IVF. Procedure: Experiment I compared the effects of SSS to human serum on mouse embryo development. Two hundred one-cell B6D2F1 mouse embryos were cultured in 100-µl droplets of human tubal fluid (HTF) containing either (1) no protein (control;n=37), (2) 15% serum from women with tubal infertility (n=44), (3) 15% serum from women with endometriosis (n=49), (4) 15% fertile donor serum (n=33), or (5) 15% SSS (n=37). Experiment II compared the effects of SSS to human serum on the development of embryos from patients undergoing IVF. Thirty-three women were included in this study. A total of 371 oocytes was cultured in HTF containing either (1) maternal or donor serum (n=140) or (2) 15% SSS (n=231). Embryo development was evaluated 48 hr after fertilization.

Results

In Experiment I, the rate of blastocyst development was evaluated at 48, 72, and 96 hr of culture. Sixty-four and nine-tenths percent of embryos cultured in SSS were morulae at 48 hr of culture (versus 5.4, 0, 8.2, and 6.1 in Groups 1, 2, 3, and 4, respectively). By 72 hr, 29.7% of these embryos had developed into blastocysts (versus 0, 0, 8.2, and 3.0, for Groups 1, 2, 3, and 4, respectively). This percentage increased to a total of 83.7 after 96 hr (versus 27.0, 20.4, 38.8, and 39.4 for Groups 1, 2, 3, and 4, respectively). Forty-three and two-tenths percent of the blastocysts cultured in SSS had hatched from their zonae by 96 hr. With the exception of Group 5, which had a rate of 9.1%, embryo hatching was not observed in any of the groups at the termination of culture (96 hr). In Experiment II there were no differences in cell stage or quality of human embryos cultured in SSS or serum, but fertilization rates tended to be better (P=0.07) for oocytes inseminated in media containing SSS (70.0%, vs 55.0% for serum).

Conclusions

SSS appears to be a superior protein source for mouse embryo growth and is as good as serum from fertile donors in promoting in vitro human embryo development. Similar content being viewed by others

References

Ogawa T, Marrs P: The effect of protein supplementation on single-cell mouse embryos. Fertil Steril 1987;47:156–616 Leung PCS, Gronow MJ, Kellow GN, Lopata A, Speirs AL, McBain JC, duPlessis YP, Johnston I: Serum supplement in human in vitro fertilization and embryo development. Fertil Steril 1984;41:36–39 Menezo Y, Testart J, Perrone D: Serum is not necessary in human in vitro fertilization, early embryo culture, and transfer. Fertil Steril 1984;42:750–755 Shirley B, Wortham JWE Jr, Witmyer J, Codon-Mahony M, Fort G: Effects of human serum and plasma on development of mouse embryos in culture media. Fertil Steril 1985;43:129–134 Dokras A, Sargent IL, Redman CWG, Balrow DH: Sera from women with unexplained infertility inhibit both mouse and human growth in vitro. Fertil Steril 1993;60:285–292 Damewood MD, Hesla JS, Schlaff WD, Hubbard M, Gearhart JD, Rock JA: Effect of serum from patients with minimal to mild endometriosis on mouse embryo development in vitro. Fertil Steril 1990;50:917–920 Abua-Musa A, Takahashi K, Okada S, Sakoda R, Kitao M: Serum from patients with threatened abortion: Effect of in vitro development of mouse embryos. J Reprod Med 1994;39:10–12 Holst N, Bertheussen K, Forsdahl F, Håkonse MB, Hansen LJ, Nielsen HI: Optimization and simplification of culture conditions in human in vitro fertilization (IVF) and preembryo replacement by serum-free media. J Vitro Fert Embryo Transfer 1990;7:47–53 Pool TB, Martin JE: High continuing pregnancy rates after in vitro fertilization-embryo transfer using medium supplemented with a plasma protein fraction containing α- and β-globulins. Fertil Steril 1994;61:714–719 Veeck LL: Atlas of the Human Oocyte and Early Conceptus, Vol. 2, Baltimore, Williams and Wilkins, 1991, p 121 Psalti I, Loumaye E, Pensis M, Depreester S, Thomas K: Evaluation of a synthetic serum substitute to replace fetal cord serum for human oocyte fertilization and embryo growth in vitro. Fertil Steril 1989;52:807–811 Author information Authors and Affiliations Rights and permissions About this article Cite this article Tucker, K.E., Hurst, B.S., Guadagnoli, S. et al. Evaluation of synthetic serum substitute versus serum as protein supplementation for mouse and human embryo culture. J Assist Reprod Genet 13, 32–37 (1996). https://doi.org/10.1007/BF02068866 Received: Accepted: Issue date: DOI: https://doi.org/10.1007/BF02068866

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MeSH descriptors

Blastocyst Fertilization in Vitro Fertilization in Vitro Plasma Substitutes Animals Blastocyst Cells, Cultured Culture Media Culture Media Female Fertility Humans Male Mice Mice, Inbred Strains Plasma Substitutes Plasma Substitutes Sperm Count Zona Pellucida Zona Pellucida

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