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. 2015 Nov 20;2015(11):CD007876.
doi: 10.1002/14651858.CD007876.pub2.

Culture media for human pre-implantation embryos in assisted reproductive technology cycles

Mohamed M A Youssef  1 , Eleni Mantikou, Madelon van Wely, Fulco Van der Veen, Hesham G Al-Inany, Sjoerd Repping, Sebastiaan Mastenbroek
Affiliations

Culture media for human pre-implantation embryos in assisted reproductive technology cycles

Mohamed M A Youssef et al. Cochrane Database Syst Rev. .

Abstract

Background: Many media are commercially available for culturing pre-implantation human embryos in assisted reproductive technology (ART) cycles. It is unknown which culture medium leads to the best success rates after ART.

Objectives: To evaluate the safety and effectiveness of different human pre-implantation embryo culture media in used for in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) cycles.

Search methods: We searched the Cochrane Menstrual Disorders and Subfertility Group's Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, the National Research Register, the Medical Research Council's Clinical Trials Register and the NHS Center for Reviews and Dissemination databases from January 1985 to March 2015. We also examined the reference lists of all known primary studies, review articles, citation lists of relevant publications and abstracts of major scientific meetings.

Selection criteria: We included all randomised controlled trials which randomised women, oocytes or embryos and compared any two commercially available culture media for human pre-implantation embryos in an IVF or ICSI programme.

Data collection and analysis: Two review authors independently selected the studies, assessed their risk of bias and extracted data. We sought additional information from the authors if necessary. We assessed the quality of the evidence using Grades of Recommendation, Assessment, Development and Evaluation (GRADE) methods. The primary review outcome was live birth or ongoing pregnancy.

Main results: We included 32 studies in this review. Seventeen studies randomised women (total 3666), three randomised cycles (total 1018) and twelve randomised oocytes (over 15,230). It was not possible to pool any of the data because each study compared different culture media.Only seven studies reported live birth or ongoing pregnancy. Four of these studies found no evidence of a difference between the media compared, for either day three or day five embryo transfer. The data from the fifth study did not appear reliable.Six studies reported clinical pregnancy rate. One of these found a difference between the media compared, suggesting that for cleavage-stage embryo transfer, Quinn's Advantage was associated with higher clinical pregnancy rates than G5 (odds ratio (OR) 1.56; 95% confidence interval (CI) 1.12 to 2.16; 692 women). This study was available only as an abstract and the quality of the evidence was low.With regards to adverse effects, three studies reported multiple pregnancies and six studies reported miscarriage. None of them found any evidence of a difference between the culture media used. None of the studies reported on the health of offspring.Most studies (22/32) failed to report their source of funding and none described their methodology in adequate detail. The overall quality of the evidence was rated as very low for nearly all comparisons, the main limitations being imprecision and poor reporting of study methods.

Authors' conclusions: An optimal embryo culture medium is important for embryonic development and subsequently the success of IVF or ICSI treatment. There has been much controversy about the most appropriate embryo culture medium. Numerous studies have been performed, but no two studies compared the same culture media and none of them found any evidence of a difference between the culture media used. We conclude that there is insufficient evidence to support or refute the use of any specific culture medium. Properly designed and executed randomised trials are necessary.

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Conflict of interest statement

None known

Figures

1
1
Study flow diagram.
2
2
Risk of bias graph: review authors' judgements about each risk of bias item presented as percentages across all included studies.
3
3
Risk of bias summary: review authors' judgements about each risk of bias item for each included study.
4
4
Forest plot of comparison: 1 Live birth or ongoing pregnancy per woman randomised, outcome: 1.1 Cleavage‐stage embryo transfer.
5
5
Forest plot of comparison: 1 Live birth or ongoing pregnancy per woman randomised, outcome: 1.2 Blastocyst‐stage embryo transfer.
6
6
Forest plot of comparison: 2 Clinical pregnancy rate per woman randomised, outcome: 2.1 Cleavage‐stage embryo transfer.
7
7
Forest plot of comparison: 2 Clinical pregnancy rate per woman randomised, outcome: 2.2 Blastocyst‐stage embryo transfer.
8
8
Forest plot of comparison: 3 Multiple pregnancy rate per woman randomised, outcome: 3.1 Cleavage‐stage embryo transfer.
9
9
Forest plot of comparison: 3 Multiple pregnancy rate per woman randomised, outcome: 3.2 Blastocyst‐stage embryo transfer.
10
10
Forest plot of comparison: 4 Miscarriage rate per woman randomised, outcome: 4.1 Cleavage‐stage embryo transfer.
11
11
Forest plot of comparison: 4 Miscarriage rate per woman randomised, outcome: 4.2 Blastocyst‐stage embryo transfer.
1.1
1.1. Analysis
Comparison 1 Live birth or ongoing pregnancy per woman randomised, Outcome 1 Cleavage‐stage embryo transfer.
1.2
1.2. Analysis
Comparison 1 Live birth or ongoing pregnancy per woman randomised, Outcome 2 Blastocyst‐stage embryo transfer.
2.1
2.1. Analysis
Comparison 2 Clinical pregnancy rate per woman randomised, Outcome 1 Cleavage‐stage embryo transfer.
2.2
2.2. Analysis
Comparison 2 Clinical pregnancy rate per woman randomised, Outcome 2 Blastocyst‐stage embryo transfer.
3.1
3.1. Analysis
Comparison 3 Multiple pregnancy rate per woman randomised, Outcome 1 Cleavage‐stage embryo transfer.
3.2
3.2. Analysis
Comparison 3 Multiple pregnancy rate per woman randomised, Outcome 2 Blastocyst‐stage embryo transfer.
4.1
4.1. Analysis
Comparison 4 Miscarriage rate per woman randomised, Outcome 1 Cleavage‐stage embryo transfer.
4.2
4.2. Analysis
Comparison 4 Miscarriage rate per woman randomised, Outcome 2 Blastocyst‐stage embryo transfer.
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References

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Kleijkers 2014 {published data only}
    1. Kleijkers SH, Montfoort AP, Smits LJ, Viechtbauer W, Roseboom TJ, Nelissen EC, et al. IVF culture medium affects post‐natal weight in humans during the first 2 years of life. Human Reproduction 2014;29(4):661‐9. - PubMed
    1. Kleijkers SH, Montfoort AP, Smits LJ, Viechtbauer W, Roseboom TJ, Nelissen EC, et al. IVF culture medium affects post‐natal weight in humans during the first 2 years of life. Human Reproduction 2014;29(4):661‐9. - PubMed
Komarovsky 2007 {published data only}
    1. Komarovsky D, Strassburger D, Bern O, Kasterstein E, Komsky E, Raziel A, et al. Comparison of two human embryo culture systems: SAGE versus Vitrolife ART line of products. Human Reproduction 2007;22(suppl 1):i167.
Kyono 1999 {published data only}
    1. Kyono K, Momma K, Chiba S, Otomo Y, Haryu M. Comparison of human tubal fluid (HTF) and P1 culture media for human IVF and ICSI. 11th World Congress on In Vitro Fertilization and Human Reproductive Genetics. 1999:202.
Kyono 2002 {published data only}
    1. Kyono K, Fukunaga N, Haigo K, Chiba S, Nakajo Y, Fuchinoue K. Two step (day 1 to 3, day 5) embryo transfers improve on‐going pregnancy rate in women with multiple failures of conception after ART. Fertility and Sterility 2002;78(3):S230.
Lane 2007 {published data only}
    1. Lane M, Gardner DK. Embryo culture medium: which is the best?. Best Practice & Research. Clinical Obstetrics & Gynaecology 2007;21:83‐100. - PubMed
Liebermann 2012 {published data only}
    1. Liebermann J, Pelts EJ, Brohammer R, Wagner Y, Sipe C, Lederer K. Single‐step vs. two‐step media: Are there any differences in outcome and implantation?. Fertility and Sterility 2012;98(3):166.
Lin 2014 {published data only}
    1. Lin YH, Hwang JL, Huang LW, Seow KM, Hsieh BC, Tzeng CR. Comparison of Quinn’s Advantage fertilization medium and tissue culture medium 199 for in vitro maturation of oocytes. Taiwanese Journal of Obstetrics & Gynecology 2014;53:17‐20. - PubMed
Lopes 2008 {published data only}
    1. Lopes VM, Pereira TR, Lopes JRC, Café TC, Almodin CG, Minguetti‐Camara VC. A comparison between GV and HTF/P1 media in IVF in‐vitro [Estudo comparativo entre meio GV e HTF/P1 em fertilizacao in‐vitro]. Jornal Brasileiro de Reproducao Assistida 2008;12:11‐4.
Mahadevan 1996 {published data only}
    1. Mahadevan MM, Miller MM, Moutos DM. Improved human zygote development in a modified Ham's F10 medium in vitro. Journal of Assisted Reproduction and Genetics 1996;13(9):722‐5. - PubMed
Mantikou 2013 {published data only}
    1. Mantikou E, Youssef MAFM, Wely M, Veen F, Al‐inany HG, Repping S, et al. Embryo culture media and IVF/ICSI success rates: A systematic review. Human Reproduction Update 2013;19:210‐20. - PubMed
Mayer 1998 {published data only}
    1. Mayer J, Weedon V, Nehchiri F, Jones E, Oehninger S, Gardner DK, et al. Blastocyst development of human embryos with poor morphological grades: a comparison of two different culture systems. Fertility and Sterility 1998;30:S118.
Mayer 1999 {published data only}
    1. Mayer IF, Nehchiri F, Kalin HL, Jones EL, Weedon VM, Lanzendorf SE, et al. Prospective randomized study of two culture media on pregnancy rates with cryo preserved zygotes. Fertility and Sterility 1999;72(3):s145.
Mayer 1999b {published data only}
    1. Mayer JF, Nehchiri F, Kalin HL, Jones EL, Weedon VM, Lanzendorf SE, et al. Prospective randomized study of two culture media on pregnancy rates with cryo preserved zygotes. Fertility and Sterility 1999;72(3):145.
Mellon 2005 {published data only}
    1. Mellon JA, Salvador CJ, Fluker MR, Yuzpe AA, Kelk DA. A randomized prospective comparison of a sequential versus single media culture system. Fertility and Sterility 2005;84(suppl 1):S449‐50.
Merry 2002 {published data only}
    1. Merry N, Attard M, Tucker K, Edgar DH. A comparison of early embryo development in two culture systems. 21st Annual Scientific Meeting of the Fertility Society of Australia. 2002:56.
Michaeli 2007 {published data only}
    1. Michaeli M, Ellenbogen A, Rotfarb N, Karchovsky E, Shalom‐Paz E, Hallack M. In vitro fertilization outcome using three types of culture media containing different concentrations of glucose for developing cleavage stage embryos. A comparative study. Fertility and Sterility 2007;88(suppl 1):S324.
Moodie 2004 {published data only}
    1. Moodie G, Schenkman E, Chao S, Sinclair P, Brenner S, Kenigsberg D. A comparison of Global, P1 and Quinn’s Advantage culture media:There was no significant difference in clinical outcome. Fertility and Sterility 2004;82(2):328.
Mortimer 2003 {published data only}
    1. Mortimer ST, Fluker MR, Yzupe AA. Effect of culture system upon fertilization rate and embryo development for patients 40 years and older. Fertility and Sterility 2003;80(1):S168.
Nelissen 2012 {published data only}
    1. Nelissen E, Moontfoort APA, Menheere PPCA, Evers JLH, Peeters LL, Dumoulin JCM. Effect of IVF culture medium on human fetal growth. Human Reproduction 2011;26(suppl 1):i67.
    1. Nelissen EC, Montfoort AP, Coonen E, Derhaag JG, Geraedts JP, Smits LJ, et al. Further evidence that culture media affect perinatal outcome: findings after transfer of fresh and cryopreserved embryos. Human Reproduction 2012;27(7):1966‐76. - PubMed
    1. Nelissen ECM, Montfoort APA, Coonen E, Derhaag JG, Evers JLH, Dumoulin JCM. In vitro culture affects the birthweight of human singletons. Human Reproduction 2010;25(suppl 1):i238.
Parinaud 1987 {published data only}
    1. Parinaud J, Monrozoies X, Reme JM, Sarramon MF, Grandjean H, Pontonnier G. Randomized trail of two media used for in vitro fertilization. European Journal of Obstetrics, Gynecology, and Reproductive Biology 1987;25:203‐8. - PubMed
Parinaud 1998 {published data only}
    1. Parinaud J, Veiga A, Vieitez G, Busquets A, Milhet P, Calderon C, et al. Randomized comparison of three media used for embryo culture after intracytoplasmic sperm injection. Human reproduction (Oxford, England) 1998;13:212‐17. - PubMed
Parinaud 1999 {published data only}
    1. Parinuard J, Milhet P, Vieitez G, Richoilley G. Use of a medium devoid of any human animal compound (SMART2) for embryo culture in intracytoplasmic sperm injection. Journal of Assisted Reproduction and Genetics 1999;16(1):13‐6. - PMC - PubMed
Perri 2001 {published data only}
    1. Perri T, Yoeli R, Mor L, Shelef M, Schwartz A, Orvieto R, et al. Culture media in assisted reproduction technology practice, a comparison study. Human Reproduction 2001;16(Suppl 1):19‐21.
Pool 2012 {published data only}
    1. Pool TB, Schoolfield J, Han D. Human embryo culture media comparisons.. Methods in Molecular Biology 2012;912:367‐86. - PubMed
Prados 2002 {published data only}
    1. Prados N, Calderon G, Crespo M, Remohi J, Caligara C, Navarro J. Culture media and protein sources in the optimisation of human embryo quality. Fertility and Sterility 2002;78(3):S245.
Quinn 1985 {published data only}
    1. Quinn P, Kerin JF, Warnes GM. Improved pregnancy rate in human in vitro fertilization with the use of a medium based on the composition of human tubal fluid. Fertility and Sterility 1985;44:493‐8. - PubMed
Quinn 1985b {published data only}
    1. Quinn P, Warnes GM, Kerin JF, Kirby C. Culture factors affecting the success rate of in vitro fertilization and embryo transfer. Annals of the New York Academy of Sciences 1985;442:195‐204. - PubMed
Quinn 1995 {published data only}
    1. Quinn P. Enhanced results in mouse and human embryo culture using a modified human tubal fluid medium lacking glucose and phosphate 1. Journal of Assisted Reproduction and Genetics 1995;12(2):97‐105. - PubMed
Quinn 1995b {published data only}
    1. Quinn P, Moinipanah R, Steinberg JM, Weathersbee PS. Successful human in vitro fertilization using a modified human tubal fluid medium lacking glucose and phosphate ions. Fertility and Sterility 1995;63:922‐4. - PubMed
Reed 2009 {published data only}
    1. Reed ML, Hamic A, Thompson DJ, Caperton CL. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study. Fertility and Sterility 2009;92(5):1783‐6. - PubMed
Rinehart 1998 {published data only}
    1. Rinehart J, Chapman C, McKiernan S, Bavister B. A protein‐free chemically defined embryo culture medium produces pregnancy rates similar to human tubal fluid (HTF) supplemented with 10% synthetic serum substitute (SSS). Human Reproduction 1998;13:59.
    1. Rinehart J, Chapman CA, McKiernan SA, Bavister B. Birth weights are similar in children from embryos cultured in a protein‐free chemically‐defined medium compared to embryos cultured in human tubal fluid (HTF) supplemented with 10% synthetic serum substitute (SSS). Fertility and Sterility 1998;70(3):S290.
Romano 2004 {published data only}
    1. Romano S, Scarselli F, Minasi MG, Martinez F, Icobelli M, Sapienza F, et al. Effect of different culture systems on embryo development potential. Human Reproduction 2004;19(suppl 1):i6‐7.
Ruiz 1999 {published data only}
    1. Ruiz A, Cobo AC, Perez‐Cano I, Herrer R, Ochoa S, Moreno D, et al. In vitro culture medium without albumin for an in vitro fertilization program. Fertility and Sterility 1999;72(suppl 1):S245‐6.
Sieren 2003 {published data only}
    1. Sieren K, Barnett B, Douglas J, Rodriguez A, Madden J, Meintjes M. Clinical outcomes of a randomized trial between G‐media versions two and three in a predominantly blastocyst program. Fertility and Sterility 2003;80(Suppl 3):S294.
Sieren 2006 {published data only}
    1. Sieren KR, Grainger DA, Tjadena BL, Balla GD. In vitro outcome of a prospective, randomized trial comparing the GIII Series™ and Quinn’s Advantage® sequential media systems for embryo development. Fertility and Sterility 2006;86:s233‐4.
Staessen 1994 {published data only}
    1. Staessen C, Abbeel E, Janssenswillen C, Devroey P, Steirteghem AC. Controlled comparison of Earle's balanced salt solution with Mėnėzo B2 medium for human in‐vitro fertilization performance. Human Reproduction 1994;10:1915‐9. - PubMed
Staessen 1998 {published data only}
    1. Staessen C, Janssenswillen C, Clerck E, Steirteghem A. Controlled comparison of commercial media for human in‐vitro fertilization: Menezo B2 medium versus Medi‐Cult universal and BM1 medium. Human Reproduction 1998;13(9):2448‐54. - PubMed
    1. Clerck E, Carle M, Henderix P, Janssens R, Laurier K, Lebrun F, et al. Comparison of Menezo B2 medium with BMI medium for human IVF. Human Reproduction 1996;11:18‐9.
Steele 2006 {published data only}
    1. Steele MG, Kopakaki A, Sriskandakumar R, Papanikou C, Sciorio R, Mary N, et al. Comparison of Medicult SequentialMedium ISM1 with non‐Sequential universalIVF Medium in an IVF/ICSI programme. Human Fertility 2006;9(4):259.
Steinkampf 2001 {published data only}
    1. Steinkampf MP, Hubbard JA, Kretzer PA, Hammond KR, Tucker M. A comparison of two sequential IVF media systems on fertilization rates and embryo quality. Fertility and Sterility 2001;76(3):S215.
Stoddart 2001 {published data only}
    1. Stoddard NR, Fleming SD. A comparison of P1/P2 and G12/G22 sequential culture media systems. 17th World Congress on Fertility and Sterility. 2001; Vol. November 2001:165.
Stoddart 2002 {published data only}
    1. Stoddart NR, Rho HC, Hussain A, Fleming SD, Bernhisel M. A comparison of the effects of short‐term oocyte culture in glucose/phosphate‐free medium or in glucose/ phosphate‐supplemented medium prior to intracytoplasmic sperm injection (ICSI) on clinical outcome. Fertility and Sterility 2002;78(1):S142‐3.
Summers‐Chase 2004 {published data only}
    1. Summers‐Chase D, Check JH, Swenson K, Yuan W, Brittingham D, Barci H. A comparison of in vitro fertilization outcome by culture media used for developing cleavage‐stage embryos. Clinical and Experimental Obstetrics & Gynecology 2004;31:179‐82. - PubMed
    1. Summers‐Chase D, Swenson K, Cochrane H, Yuan W, Brittingham D, Check JH. A random comparison of a new non‐sequential media, a new sequential media, and a standard non‐sequential media on in vitro fertilization outcome. Fertility and Sterility 2002;78:S287.
Sun 2001 {published data only}
    1. Sun M, Feng Y, Gao Q. Application of sequential culture in assisted reproductive technique. Zhonghua Fu Chan Ke Za Zhi 2001;36(2):92‐4. - PubMed
Szell 1998 {published data only}
    1. Szell AZ, Antaran JM, Chetkowski RJ. Pregnancy and implantation rates from the transfer of human embryos cultured in P1 or human tubal fluid medium and transferred with or without assisted hatching. Fertility and Sterility 1998;30:S495.
Tekpetey 1998 {published data only}
    1. Tekpetey FR, Martin JSB, Tummon IS, Nisker J, Power S. Randomised control comparison of human tubal fluid (HTF) and P1 culture media for human IVF. Fertility and Sterility 1998;70:S17.
Thorsell 2003 {published data only}
    1. Thorsell LP, Iwaszko MA, Palmer MA, Hook R, Dorfmann AD, Blauer KL. Use of two commercially available sequential media culture systems does not affect pregnancy rates. Fertility and Sterility 2003;80(3):296‐7.
Torres 2012 {published data only}
    1. Torres P, Peinado I, Orden M, Monzo A, Rubio JM, Pellicer A. Evaluation of the efficiency of different commercial systems of embryo culture [Evaluacion de la eficacia de diferentes sistemas comerciales de cultivo embrionario]. Revista Iberoamericana de Fertilidad y Reproduccion Humana 2012;27(5):417‐24.
Utsunomiya 2002 {published data only}
    1. Utsunomiya T, Naitu T, Nagaki M. A prospective trial of blastocyst culture and transfer. Human Reproduction 2002;17:1846‐51. - PubMed
Van der Auwera 2001 {published data only}
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Van Langenendonckt 2001 {published data only}
    1. Langendonckt A, Godin PA, Demylle D, Beliard A, Wyns C. Pregnancy outcome following human embryo culture in G11/G22 and Sydney IVF cleavage/blastocyst sequential media: a prospective randomized comparative study. Human Reproduction 2000;15:141‐2.
    1. Langenendonckt 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. Fertility and Sterility 2001;76:1023‐31. - PubMed
Vansteenburgge 2007 {published data only}
    1. Vansteenbrugge A, Vastersaegher C, Fontanie P, Klay C, Rodez D, Pauwels PC. Comparison of a single medium with sequential media for the development of human zygotes to the blastocyst stage. Fertility and Sterility 2007;88(suppl 1):S319.
Verheyen 2003 {published data only}
    1. Verheyen G, Landuyt L, Joris H, Devroey P, Steirtegham A. Different embryo development patterns in different sequential media appear to result in similar pregnancy rates. Human Reproduction 2003;18(Suppl 1):xviii36‐8.
Verza 2003 {published data only}
    1. Verza S, Schneider DT, Esteves SC. Comparison of two systems to culture human embryos up to day 3: a prospective randomized study. Fertility and Sterility 2003;88(suppl 1):149‐50.
Warnes 1997 {published data only}
    1. Warnes GM, Payne D, Jerrrey R, Hourigan L, Kirby C, Kerin J, et al. Reduced pregnancy rates following the transfer of human embryos frozen or thawed in culture media supplemented with normal serum albumin. Human Reproduction 1997;12(7):1525‐30. - PubMed
Westin 2003 {published data only}
    1. Westin C, Borg K, Hillensjo T, Jakobsson AH, Olofsson J, Reismer E, et al. A comparison between a conventional medium and a new generation of sequential media (GIII) for transfer of early cleavage embryo. Human Reproduction 2003;18(1):151.
Wirleitner 2010 {published data only}
    1. Wirleitner B, Vanderzwalmen P, Stecher A, Zech MH, Zintz M, Zech NH. Individual demands of human embryos on IVF culture medium: influence on blastocyst development and pregnancy outcome. Reproductive Biomedicine Online 2010;21(6):776‐82. - PubMed
Xella 2009 {published data only}
    1. Xella S, Marsella T, Tagliasacchi D, Giulini S, Marca A, Tirelli A, et al. Embryo quality and implantation rate in two different culture media: ISM1 versus Universal IVF Medium. Fertility and Sterility 2009;93(6):1859‐63. - PubMed
Zech 2006 {published data only}
    1. Zech N, Stecher A, Zech H, Uher P, Vanderzwalmen P. Prospective analysis of embryo development to day 5 and transfer outcome in sequential medium (G1.3‐G2.3) vs. a one step protocol (Global medium). Human Reproduction 2006;21(suppl 1):i162‐3.
Zhang 2002 {published data only}
    1. Zhang X, Horan K, Kallmann K, Confino E, Kazer RR. A comparison between culture media from two commercial vendors for their ability to support embryo development. Human Reproduction 2002;17(suppl 1):137.

References to ongoing studies

Dal Canto 2013 {published and unpublished data}
    1. Dal Canto, et al. ISM1 Versus EmbrioGen Media for Embryo Culture With Previous Failure Implantation in ART Treatments. http://clinicaltrials.gov/ct2/show/NCT01689454?term=ivf+culture&rank=5.
Hardarson 2014 {published data only}
    1. Harsarson, et al. Comparison of Human Embryo Development Using Sequential and Single Media With Time‐lapse Imaging. http://www.clinicaltrials.gov/ct2/show/record/NCT01939626?term=ivf+cultu....
Mastenbroek 2013 {published data only}
    1. Mastenbroek, et al. A multi‐center double‐blind randomized controlled trial of two media for the in vitro culture of human preimplantation embryos.. http://www.trialregister.nl/trialreg/admin/rctview.asp?TC=1979.
Werner 2014 {published data only}
    1. Werner MD, Hong KH, Franasiak JM, Reda CV, et al. Sequential (S) vs. monophasic (M) media impact trial (Summit): A paired randomized controlled trial comparing blastulation rates or aneuploidy risk amongst sibling zygotes.. Fertility and Sterility. 2014; Vol. 102(3 SUPPL 1):e129‐30. - PubMed
    1. Werner, et al. Sequential vs. Monophasic Media Impact Trial (SuMMIT). http://www.clinicaltrials.gov/ct2/show/record/NCT01917240?term=ivf+cultu.... - PubMed

Additional references

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Doherty 2000
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Fauque 2007
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Gardner 2002 a
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Gardner 2002 b
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Higgins 2011
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Hoelker 2010
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Khosla 2001
    1. Khosla S, Dean W, Reik W, Feil R. Culture of preimplantation embryos and its long‐term effects on gene expression and phenotype. Human Reproduction Update 2001;7(4):419‐27. - PubMed
Menezo 2004
    1. Menezo Y, Testart J, Perrone D. Serum is not necessary in human invitro fertilization, early embryo culture, and transfer. Fertility and Sterility 1984;42:750‐5. - PubMed
Zander 2006
    1. Zander DL, Thompson JG, Lane M. Perturbations in mouse embryo development and viability caused by ammonium are more severe after exposure at the cleavage stages. Biology of Reproduction 2006;74(2):288‐94. - PubMed

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