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• W2296186698 abstract "With increased use of comprehensive chromosome screening (CCS), the question remains as to why some practices do not experience the same high levels of clinical success after implementation of the approach. Indeed, the debate surrounding the efficacy and usefulness of blastocyst biopsy and CCS continues. Importantly, several variables impact the success of an assisted reproductive technology cycle. Transfer of a euploid embryo is but one factor in an intricate system that requires numerous steps to occur successfully. Certainly, the culture environment and the manipulations of the embryo during its time in the laboratory can impact its reproductive potential. Environmental stressors ranging from culture media to culture conditions and even culture platform can impact biochemical, metabolic, and epigenetic patterns that can affect the developing cell independent of chromosome number. Furthermore, accompanying procedures, such as biopsy and vitrification, are complex and, when performed improperly, can negatively impact embryo quality. These are areas that likely still carry room for improvement within the IVF laboratory. With increased use of comprehensive chromosome screening (CCS), the question remains as to why some practices do not experience the same high levels of clinical success after implementation of the approach. Indeed, the debate surrounding the efficacy and usefulness of blastocyst biopsy and CCS continues. Importantly, several variables impact the success of an assisted reproductive technology cycle. Transfer of a euploid embryo is but one factor in an intricate system that requires numerous steps to occur successfully. Certainly, the culture environment and the manipulations of the embryo during its time in the laboratory can impact its reproductive potential. Environmental stressors ranging from culture media to culture conditions and even culture platform can impact biochemical, metabolic, and epigenetic patterns that can affect the developing cell independent of chromosome number. Furthermore, accompanying procedures, such as biopsy and vitrification, are complex and, when performed improperly, can negatively impact embryo quality. These are areas that likely still carry room for improvement within the IVF laboratory. Discuss: You can discuss this article with its authors and with other ASRM members at http://fertstertforum.com/swainj-optimizing-ivf-lab-conditions/ Discuss: You can discuss this article with its authors and with other ASRM members at http://fertstertforum.com/swainj-optimizing-ivf-lab-conditions/ Since the early days of embryo culture in vitro, various modifications to the culture system have been explored in attempts to optimize embryo development and increase the number of good-quality embryos available for transfer. This has resulted in most labs being able to grow numerous high-quality embryos, often to the blastocyst stage, and has helped lead to improved overall pregnancy rates. In turn, embryo selection methods have become important as IVF centers strive to further increase efficacy, promote single ETs, and reduce time to pregnancy and live birth. Toward this end, comprehensive chromosome screening (CCS), or preimplantation genetic screening (PGS) examining embryos for all 23 pairs of chromosomes, has received an immense amount of attention. However, this approach is controversial, and the ability to improve outcomes is questioned by some investigators. Of note, different laboratories seem to experience differing levels of success when applying CCS. This may suggest that laboratory conditions could be impacting efficacy. Despite the significant impact of gamete quality on subsequent embryo development, it is often assumed that suboptimal embryo culture conditions are largely responsible for poor embryo development in vitro. While embryo culture conditions may not be able to overcome the inherent limitations imposed by the sperm or egg, the quest to improve the culture environment continues. Similarly, optimizing other procedures within the IVF laboratory to minimize the stress imposed on the embryo is an ongoing endeavor. The IVF laboratory must not only grow competent embryos but must ensure this competency is maintained after various manipulations, such as biopsy and cryopreservation. Culture media have vastly improved since the first attempt at IVF, when simple somatic cell media were used with serum supplementation. Laboratories often made their own media, which introduced variables that could impact efficacy. Various modifications were made, often using animal embryos as model systems, and eventually media were formulated specifically for human embryos (1Quinn P. Kerin J. Warnes G. Improved pregnancy rate in human in vitro fertilization with the use of a medium based on the composition of human tubal fluid.Fertil Steril. 1985; 44: 493-498Abstract Full Text PDF PubMed Google Scholar, 2Menezo Y. Testart J. Perrone D. Serum is not necessary in human in vitro fertilization, early embryo culture, and transfer.Fertil Steril. 1984; 42: 750-755Abstract Full Text PDF PubMed Google Scholar). The introduction of commercially produced media vastly improved consistency, resulted in the ability to culture embryos to the blastocyst stage, and undoubtedly aided in the success of IVF. While numerous studies have explored the impact on embryo quality of altering the energy substrate composition, supplementing macromolecules, adding growth factors, and modifying other media constituents and culture conditions, with technological improvements and new approaches to assess embryo metabolism, morphokinetics, and other means of viability assessment, there remains the possibility that media formulations may still be further refined and improved to benefit embryo development and improve resulting clinical outcomes. Various studies have attempted to compare the efficacy of various embryo culture media with the goal of determining whether one is superior to another for the growth of human embryos (3Balaban B. Urman B. Comparison of two sequential media for culturing cleavage-stage embryos and blastocysts: embryo characteristics and clinical outcome.Reprod Biomed Online. 2005; 10: 485-491Abstract Full Text PDF PubMed Google Scholar, 4Ben-Yosef D. Amit A. Azem F. Schwartz T. Cohen T. Mei-Raz N. et al.Prospective randomized comparison of two embryo culture systems: P1 medium by Irvine Scientific and the Cook IVF.Medium J Assist Reprod Genet. 2004; 21: 291-295Crossref PubMed Scopus (0) Google Scholar, 5Mantikou E. Youssef M.A. van Wely M. van der Veen F. Al-Inany H.G. Repping S. et al.Embryo culture media and IVF/ICSI success rates: a systematic review.Hum Reprod Update. 2013; 19: 210-220Crossref PubMed Scopus (47) Google Scholar, 6Mauri A.L. Petersen C.G. Baruffi R.L. Franco Jr., J.G. A prospective, randomized comparison of two commercial media for ICSI and embryo culture.J Assist Reprod Genet. 2001; 18: 378-381Crossref PubMed Scopus (0) Google Scholar, 7Parinaud J. Veiga A. Vieitez G. Busquets A. Milhet P. Calderon G. et al.Randomized comparison of three media used for embryo culture after intracytoplasmic sperm injection.Hum Reprod. 1998; 13: 212-217Crossref PubMed Google Scholar, 8Van Langendonckt A. Demylle D. Wyns C. Nisolle M. Donnez J. Comparison of G1.2/G2.2 and Sydney IVF cleavage/blastocyst sequential media for the culture of human embryos: a prospective, randomized, comparative study.Fertil Steril. 2001; 76: 1023-1031Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar, 9Zollner K.P. Zollner U. Schneider M. Dietl J. Steck T. Comparison of two media for sequential culture after IVF and ICSI shows no differences in pregnancy rates: a randomized trial.Med Sci Monitor. 2004; 10: CR1-CR7PubMed Google Scholar, 10Reed M.L. Hamic A. Thompson D.J. Caperton C.L. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study.Fertil Steril. 2009; 92: 1783-1786Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar, 11Xella S. Marsella T. Tagliasacchi D. Giulini S. La Marca A. Tirelli A. et al.Embryo quality and implantation rate in two different culture media: ISM1 versus Universal IVF Medium.Fertil Steril. 2010; 93: 1859-1863Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar, 12Summers M.C. Bird S. Mirzai F.M. Thornhill A. Biggers J.D. Human preimplantation embryo development in vitro: a morphological assessment of sibling zygotes cultured in a single medium or in sequential media.Hum Fertil (Camb). 2013; 16: 278-285Crossref PubMed Scopus (0) Google Scholar). Unfortunately, many of these studies are underpowered, do not use proper experimental design, and fail to control for critical variables that affect culture media performance (13Pool T.B. Schoolfield J. Han D. Human embryo culture media comparisons.Methods Mol Biol. 2012; 912: 367-386PubMed Google Scholar). Thus, drawing concrete conclusions about the superiority of any individual commercially available culture medium is difficult. Furthermore, no study has compared all available media in a controlled fashion, making it impossible to declare an “optimal” recipe. A recent systematic review on randomized control trials (RCTs) concluded that a conventional meta-analysis comparing embryo culture media was not possible (5Mantikou E. Youssef M.A. van Wely M. van der Veen F. Al-Inany H.G. Repping S. et al.Embryo culture media and IVF/ICSI success rates: a systematic review.Hum Reprod Update. 2013; 19: 210-220Crossref PubMed Scopus (47) Google Scholar). Only four trials reported on live birth (9Zollner K.P. Zollner U. Schneider M. Dietl J. Steck T. Comparison of two media for sequential culture after IVF and ICSI shows no differences in pregnancy rates: a randomized trial.Med Sci Monitor. 2004; 10: CR1-CR7PubMed Google Scholar, 14Campo R. Binda M.M. Van Kerkhoven G. Frederickx V. Serneels A. Roziers P. et al.Critical reappraisal of embryo quality as a predictive parameter for pregnancy outcome: a pilot study.Facts Views Vision ObGyn. 2010; 2: 289-295Google Scholar, 15Paternot G. Debrock S. D’Hooghe T.M. Spiessens C. Early embryo development in a sequential versus single medium: a randomized study.Reprod Biol Endocrinol. 2010; 8: 83Crossref PubMed Scopus (0) Google Scholar, 16Nelissen E.C. Van Montfoort A.P. Coonen E. Derhaag J.G. Geraedts J.P. Smits L.J. et al.Further evidence that culture media affect perinatal outcome: findings after transfer of fresh and cryopreserved embryos.Hum Reprod. 2012; 27: 1966-1976Crossref PubMed Scopus (0) Google Scholar), and one reported a significant difference. Nine trials reported ongoing and/or clinical pregnancy rates, of which only four showed a significant difference (16Nelissen E.C. Van Montfoort A.P. Coonen E. Derhaag J.G. Geraedts J.P. Smits L.J. et al.Further evidence that culture media affect perinatal outcome: findings after transfer of fresh and cryopreserved embryos.Hum Reprod. 2012; 27: 1966-1976Crossref PubMed Scopus (0) Google Scholar). Pooling the data did not reveal a superior culture medium. A follow-up review found similar results. Thus, it is unknown what culture medium leads to the best success rates in IVF/intracytoplasmic sperm injection (ICSI) (5Mantikou E. Youssef M.A. van Wely M. van der Veen F. Al-Inany H.G. Repping S. et al.Embryo culture media and IVF/ICSI success rates: a systematic review.Hum Reprod Update. 2013; 19: 210-220Crossref PubMed Scopus (47) Google Scholar, 17Youssef M.M. Mantikou E. van Wely M. Van der Veen F. Al-Inany H.G. Repping S. et al.Culture media for human pre-implantation embryos in assisted reproductive technology cycles.Cochrane Database Syst Rev. 2015; : CD007876PubMed Google Scholar). Currently, human preimplantation embryo culture media are broadly grouped into single-step or sequential media, with the former lending itself to interrupted or uninterrupted culture. Several reviews are available describing the evolution of these media systems and the potential benefits of either approach (18Lane M. Gardner D.K. Embryo culture medium: which is the best?.Best Pract Res Clin Obstet Gynaecol. 2007; 21: 83-100Abstract Full Text Full Text PDF PubMed Scopus (94) Google Scholar, 19Pool T.B. An update on embryo culture for human assisted reproductive technology: media, performance, and safety.Semin Reprod Med. 2005; 23: 309-318Crossref PubMed Scopus (0) Google Scholar, 20Pool T.B. Development of culture media for human assisted reproductive technology.Fertil Steril. 2004; 81: 287-289Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar, 21Biggers J.D. Summers M.C. Choosing a culture medium: making informed choices.Fertil Steril. 2008; 90: 473-483Abstract Full Text Full Text PDF PubMed Scopus (68) Google Scholar, 22Machtinger R. Racowsky C. Culture systems: single step.Methods Mol Biol. 2012; 912: 199-209PubMed Google Scholar, 23Quinn P. Culture systems: sequential.Methods Mol Biol. 2012; 912: 211-230PubMed Google Scholar). Unfortunately, few well-designed, prospective RCTs exist that compare single-step media to sequential media, and it is impossible to say which approach is superior (Table 1, Table 2 ). Most culture media within these two broad categories are similar in terms of composition. They contain glucose, pyruvate, and lactate at varying concentrations to permit embryo development past traditional developmental blocks. The majority of culture media also contain some assortment of amino acids. Because commercial embryo culture companies do not publish concentrations of media components, it is difficult to discern why one culture media might be superior to another, although methods exist to approximate media composition (Table 3) (27Morbeck D.E. Krisher R.L. Herrick J.R. Baumann N.A. Matern D. Moyer T. Composition of commercial media used for human embryo culture.Fertil Steril. 2014; 102: 759-766.e9Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar).Table 1Prospective randomized trials comparing sequential and single-step media using patient randomization.StudyMedia comparedTotal transfersTransfer day (2/3 vs. 5/6)Clinical pregnancy rate/transfer (sequential vs. single)Total implantation rateDay 3 implantation rate (sequential vs. single)Day 5/6 implantation rate (sequential vs. single)15Paternot G. Debrock S. D’Hooghe T.M. Spiessens C. Early embryo development in a sequential versus single medium: a randomized study.Reprod Biol Endocrinol. 2010; 8: 83Crossref PubMed Scopus (0) Google ScholarSydney IVF cleavage vs. GM50170 vs. 77Day 3 or day 520% vs. 28% (P<.032)19.6% vs. 26%20% vs. 43%24Sepulveda S. Garcia J. Arriaga E. Diaz J. Noriega-Portella L. Noriega-Hoces L. In vitro development and pregnancy outcomes for human embryos cultured in either a single medium or in a sequential media system.Fertil Steril. 2009; 91: 1765-1770Abstract Full Text Full Text PDF PubMed Scopus (45) Google ScholarECM/multiblast vs. Global38 vs. 40Day 5/652.6% vs. 72.5%36.8% vs. 57.5% (P<.08)36.8% vs. 57.5% (P<.008)14Campo R. Binda M.M. Van Kerkhoven G. Frederickx V. Serneels A. Roziers P. et al.Critical reappraisal of embryo quality as a predictive parameter for pregnancy outcome: a pilot study.Facts Views Vision ObGyn. 2010; 2: 289-295Google ScholarISM1 vs. GM50183 vs. 84Day 2/326.5% vs. 22.6%17.8% vs. 20.0%17.8% vs. 20.0%Note: Retrospective analyses or preliminary studies were not included. (For the purpose of this comparison, media that were formulated to only support embryo development through day 3 of development, such as P1, IVF-50, or human tubal fluid, are not viewed as single-step media. Single-step media are media formulated to support embryo development throughout development to the blastocyst stage). Paternot et al. 5Mantikou E. Youssef M.A. van Wely M. van der Veen F. Al-Inany H.G. Repping S. et al.Embryo culture media and IVF/ICSI success rates: a systematic review.Hum Reprod Update. 2013; 19: 210-220Crossref PubMed Scopus (47) Google Scholar used different handling media and fertilization media between groups. Open table in a new tab Table 2Prospective randomized trials comparing sequential and single-step media using sibling embryo splits.StudyMedia comparedNo. of patients (total transfers)No. of transfers for each mediaTransfer day (3 vs. 4/5)Day 3 clinicial pregnancy rate (sequential vs. single)Day 4/5 clinicial pregnancy rate (sequential vs. single)Day 3 implantation rate (sequential vs. single)Day 4/5 implantation rate (sequential vs. single)10Reed M.L. Hamic A. Thompson D.J. Caperton C.L. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study.Fertil Steril. 2009; 92: 1783-1786Abstract Full Text Full Text PDF PubMed Scopus (37) Google ScholarG5 Series vs. Global807 vs. 21 (52 had embryos from both)Day 3 or day 525Ciray H.N. Aksoy T. Goktas C. Ozturk B. Bahceci M. Time-lapse evaluation of human embryo development in single versus sequential culture media—a sibling oocyte study.J Assist Reprod Genet. 2012; 29: 891-900Crossref PubMed Scopus (56) Google ScholarECM/multiblast vs. SSM4913 vs. 17 (19 had embryos from both)Days 3, 4, and 50% vs. 40% (0/1 vs. 2/5)66.6% vs. 58.3%0% vs. 28.6%57.1% vs. 52.6%26Basile N. Morbeck D. Garcia-Velasco J. Bronet F. Meseguer M. Type of culture media does not affect embryo kinetics: a time-lapse analysis of sibling oocytes.Hum Reprod. 2013; 28: 634-641Crossref PubMed Scopus (39) Google ScholarSage vs. Global7317 vs. 25 (31 had embryos from both)Day 358.8% vs. 48%38.2% vs. 42%Note: Retrospective analyses or preliminary studies were not included. (For the purpose of this comparison, media that were formulated to only support embryo development through day 3 of development, such as P1, IVF-50 or human tubal fluid, are not viewed as single step. Reed et al. 10Reed M.L. Hamic A. Thompson D.J. Caperton C.L. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study.Fertil Steril. 2009; 92: 1783-1786Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar single-step media did not have any media refreshment during day 5 culture. (CPR and implantation breakdown data for day 3 vs. day 5 from Reed et al. [10Reed M.L. Hamic A. Thompson D.J. Caperton C.L. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study.Fertil Steril. 2009; 92: 1783-1786Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar] is not available). Open table in a new tab Table 3Concentrations of essential media components from commercial embryo culture media as determined via various analytical methods.VariableIVFOnline globalIrvine CSCVitrolifeSageCookIn vitro careOrigioG1G2QACMQABMSICMSIBMIVC1IVC3ISM1BAGlucose (mM)0.20.50.53.40.12.80.33.102.71.01.0Citrate (mM)00.010.080.0800.160000.160.020.003Lactate (mM)4.85.610.86.03.93.91.81.810.19.43.22.4Pyruvate (mM)0.200.170.300.070.520.070.360.310.080.092.00.17L:P ratio243336867.5565.05.912610518.51.2Amino acid concentrations (μM) EssentialArg32828103600313252520590138124Cys5246054054232094238His1111050121010288601889954Ile22120202490209171690388147208Leu23021402650227181820408158217Lys23222302600223181740417148179Met515306305644301008954Phe11210601250106886020090104Thr2161950242021018172037481211Trp282603002822205110021Tyr10095011401001211401867091Val23321502560224171790428356225 NonessentialAla6562148151001351350136338124Asn52571261291121248884011373104Asp5547009310481850956578Glu54460000908701031102Gln0000003026007780Gly63581351411191316647481501211760701Pro51481121139310385800998296Ser58551271301071239289010996113Tau00131012212064896380002960Note: QACM = Quinn's Advantage Cleavage media; QABM = Quinn's Advantage Blastocyst media; SICM = Sydney IVF Cleavage media; SIBM = Sydney IVF Blastocyst media; BA = Blastassist. (Adapted from Morbeck et al. [27Morbeck D.E. Krisher R.L. Herrick J.R. Baumann N.A. Matern D. Moyer T. Composition of commercial media used for human embryo culture.Fertil Steril. 2014; 102: 759-766.e9Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar]). Open table in a new tab Note: Retrospective analyses or preliminary studies were not included. (For the purpose of this comparison, media that were formulated to only support embryo development through day 3 of development, such as P1, IVF-50, or human tubal fluid, are not viewed as single-step media. Single-step media are media formulated to support embryo development throughout development to the blastocyst stage). Paternot et al. 5Mantikou E. Youssef M.A. van Wely M. van der Veen F. Al-Inany H.G. Repping S. et al.Embryo culture media and IVF/ICSI success rates: a systematic review.Hum Reprod Update. 2013; 19: 210-220Crossref PubMed Scopus (47) Google Scholar used different handling media and fertilization media between groups. Note: Retrospective analyses or preliminary studies were not included. (For the purpose of this comparison, media that were formulated to only support embryo development through day 3 of development, such as P1, IVF-50 or human tubal fluid, are not viewed as single step. Reed et al. 10Reed M.L. Hamic A. Thompson D.J. Caperton C.L. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study.Fertil Steril. 2009; 92: 1783-1786Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar single-step media did not have any media refreshment during day 5 culture. (CPR and implantation breakdown data for day 3 vs. day 5 from Reed et al. [10Reed M.L. Hamic A. Thompson D.J. Caperton C.L. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study.Fertil Steril. 2009; 92: 1783-1786Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar] is not available). Note: QACM = Quinn's Advantage Cleavage media; QABM = Quinn's Advantage Blastocyst media; SICM = Sydney IVF Cleavage media; SIBM = Sydney IVF Blastocyst media; BA = Blastassist. (Adapted from Morbeck et al. [27Morbeck D.E. Krisher R.L. Herrick J.R. Baumann N.A. Matern D. Moyer T. Composition of commercial media used for human embryo culture.Fertil Steril. 2014; 102: 759-766.e9Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar]). Recently, several single-step media have been introduced to be compatible with uninterrupted culture and use with time-lapse technology. Whether any observed improvements in outcomes are due to the media composition itself, to less handling and stress to the cells for routine observation, or to the newer incubator design or gas concentrations is unknown. That being said, composition of culture media is still important in trying to improve current culture conditions. The balance of organic and inorganic salts is crucial (28Baltz J.M. Media composition: salts and osmolality.Methods Mol Biol. 2012; 912: 61-80PubMed Google Scholar). The composition and ratios of energy substrates is also critical. One area that likely deserves extra attention in optimizing media performance is the composition of amino acids. Amino acids support numerous cellular processes, including acting as metabolites, osmolytes, antioxidants, and buffers, and likely help alleviate stress in the embryo culture system (29Lane M. Mechanisms for managing cellular and homeostatic stress in vitro.Theriogenology. 2001; 55: 225-236Abstract Full Text PDF PubMed Scopus (0) Google Scholar). Even a brief period of culture with no amino acids can impair mouse embryo development (30Gardner D.K. Lane M. Alleviation of the “2-cell block” and development to the blastocyst of CF1 mouse embryos: role of amino acids, EDTA and physical parameters.Hum Reprod. 1996; 11: 2703-2712Crossref PubMed Scopus (0) Google Scholar). Thus, all embryo culture media should contain some assortment of amino acids. While animal studies have given some insight into the positive and negative effects of individual and combinations of amino acids (31McKiernan S.H. Clayton M.K. Bavister B.D. Analysis of stimulatory and inhibitory amino acids for development of hamster one-cell embryos in vitro.Mol Reprod Dev. 1995; 42: 188-199Crossref PubMed Scopus (0) Google Scholar, 32Lane M. Gardner D.K. Differential regulation of mouse embryo development and viability by amino acids.J Reprod Fertil. 1997; 109: 153-164Crossref PubMed Google Scholar, 33Lane M. Hooper K. Gardner D.K. Effect of essential amino acids on mouse embryo viability and ammonium production.J Assist Reprod Genet. 2001; 18: 519-525Crossref PubMed Scopus (0) Google Scholar), detailed assessments of individual amino acids and all possible combinations and ratios have not been conducted. Additionally, while controversial, a concern with the inclusion of amino acids in media is the buildup of ammonium and its potential negative impact on embryo and fetal development (34Biggers J.D. McGinnis L.K. Summers M.C. Discrepancies between the effects of glutamine in cultures of preimplantation mouse embryos.Reprod Biomed Online. 2004; 9: 70-73Abstract Full Text PDF PubMed Google Scholar). This potential problem may be alleviated through media change in a sequential system. However, use of proper assortment and concentrations of amino acids and culture conditions is also a feasible approach. Modern embryo culture media should include the dipeptide form of glutamine to reduce ammonium production (35Biggers J.D. McGinnis L.K. Lawitts J.A. Enhanced effect of glycyl-L-glutamine on mouse preimplantation embryos in vitro.Reprod Biomed Online. 2004; 9: 59-69Abstract Full Text PDF PubMed Google Scholar, 36Summers M.C. McGinnis L.K. Lawitts J.A. Biggers J.D. Mouse embryo development following IVF in media containing either L-glutamine or glycyl-L-glutamine.Hum Reprod. 2005; 20: 1364-1371Crossref PubMed Scopus (0) Google Scholar, 37Lane M. Gardner D.K. Ammonium induces aberrant blastocyst differentiation, metabolism, pH regulation, gene expression and subsequently alters fetal development in the mouse.Biol Reprod. 2003; 69: 1109-1117Crossref PubMed Scopus (0) Google Scholar, 38Virant-Klun I. Tomazevic T. Vrtacnik-Bokal E. Vogler A. Krsnik M. Meden-Vrtovec H. Increased ammonium in culture medium reduces the development of human embryos to the blastocyst stage.Fertil Steril. 2006; 85: 526-528Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar). Whether the use of other dipeptide amino acids could further improve the culture environment is unknown (39Moravek M. Fisseha S. Swain J.E. Dipeptide forms of glycine support mouse preimplantation embryo development in vitro and provide protection against high media osmolality.J Assist Reprod Genet. 2012; 29: 283-290Crossref PubMed Scopus (0) Google Scholar), although a clearer understanding of embryo metabolism and use of dipeptides is likely warranted (39Moravek M. Fisseha S. Swain J.E. Dipeptide forms of glycine support mouse preimplantation embryo development in vitro and provide protection against high media osmolality.J Assist Reprod Genet. 2012; 29: 283-290Crossref PubMed Scopus (0) Google Scholar, 40Swain J. Dipeptide glutamine functions as an organic osmolyte in mouse preimplantations embryos but does not support blastocyst hatching at the same rate as individual component amino acids.Fertil Steril. 2015; 104: e313Abstract Full Text Full Text PDF Google Scholar). Macromolecule supplementation is another area that may lend itself to media improvement. Primary protein supplements used in clinical embryo culture media include human serum albumin (HSA), as well as complex protein supplements containing HSA and a combination of alpha and beta globulins (41Weathersbee P.S. Pool T.B. Ord T. Synthetic serum substitute (SSS): a globulin-enriched protein supplement for human embryo culture.J Assist Reprod Genet. 1995; 12: 354-360Crossref PubMed Scopus (0) Google Scholar, 42Pool T.B. Martin J.E. High continuing pregnancy rates after in vitro fertilization–embryo transfer using medium supplemented with a plasma protein fraction containing alpha- and beta-globulins.Fertil Steril. 1994; 61: 714-719Abstract Full Text PDF PubMed Google Scholar). Macromolecules can act as a surfactant and as a nitrogen source, stabilize membranes and modulate the physical microenvironment, act as a carrier molecule for other compounds, or even bind trace elements and toxins. Unfortunately, macromolecule/protein supplements are one of the least defined components of the culture system and thus are a large source of potential variation. For example, stabilizers/preservatives that are used when making protein solutions, such as octanoic acid, vary in concentration and may be embryotoxic (43Leonard P.H. Charlesworth M.C. Benson L. Walker D.L. Fredrickson J.R. Morbeck D.E. Variability in protein quality used for embryo culture: embryotoxicity of the stabilizer octanoic acid.Fertil Steril. 2013; 100: 544-549Abstract Full Text Full Text PDF PubMed Google Scholar, 44Fredrickson J. Krisher R. Morbeck D.E. The impact of the protein stabilizer octanoic acid on embryonic development and fetal growth in a murine model.J Assist Reprod Genet. 2015; 32: 1517-1524Crossref PubMed Google Scholar). Additionally, other proteins, growth factors, and hormone “contaminants” may be present at varying levels in protein preparations that are supposed to be predominately albumin (45Morbeck D.E. Paczkowski M. Fredrickson J.R. Krisher R.L. Hoff H.S. Baumann N.A. et al.Composition of protein supplements used for human embryo culture.J Assist Reprod Genet. 2014; 31: 1703-1711Crossref PubMed Scopus (0) Google Scholar, 46Meintjes M. Media composition: macromolecules and embryo growth.Methods Mol Biol. 2012; 912: 107-127PubMed Google Scholar, 47Dyrlund T." @default.
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• W2296186698 date "2016-03-01" @default.
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• W2296186698 title "Optimizing the culture environment and embryo manipulation to help maintain embryo developmental potential" @default.
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