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Embryo culture

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to the blastocyst stage, one medium may be used for culture to day 3, and a second medium is used for culture thereafter. Single or sequential medium are equally effective for the culture of human embryos to the blastocyst stage. Artificial embryo culture media basically contain glucose, pyruvate, and energy-providing components, but the addition of amino acids, nucleotides, vitamins, and cholesterol improve the performance of embryonic growth and development. Specifically, embryo culture media contain more pyruvate concentration than glucose in the cleavage phase and more glucose concentration than pyruvato in the blastocyst phase. This is because before day 3 the embryo uses the oocyte reserves, however, from day 3 to the blastocyst it starts to express different proteins to continue its development, so it starts to degrade glucose (it needs more glucose in this case). Also substances like antioxidants, antibiotics, macromolecules, hormones and growth factors can be added. Methods to permit dynamic embryo culture with fluid flow and embryo movement are also available. A new method in development uses the uterus as an incubator and the naturally occurring intrauterine fluids as culture medium by encapsulating the embryos in a permeable intrauterine vessel.
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can be used, in which the embryo is sequentially placed in different media, with different formulations based on the different concentration and composition of the tubal and uterine fluid in relation to change in the metabolic activity of the embryo during its development. For example, when culturing
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Regarding the incubator, technicians should place one patient per incubator and avoid frequent door opening. Taking into account the number of embryos used in the culture, group embryo culture is recommended, so they can exchange growing factors while time is saved in the lab but embryo fusion is a
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While it has been hypothesized that incubating at a temperature lower than 37 °C may be a more accurate recreation of the temperature in the female reproductive tract, the evidence is uncertain whether different temperatures for embryo culture have different effects on pregnancy or live birth
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The first thing to take into account are the oxygen and carbon dioxide conditions because they must be as similar as the uterus ones as possible. It is for this reason that oxygen has to be at 5% and carbon dioxide at 6% (depending on altitude). On the other hand, temperature must be set at 37
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Blockeel, C.; Mock, P.; Verheyen, G.; Bouche, N.; Le Goff, P.; Heyman, Y.; Wrenzycki, C.; Höffmann, K.; Niemann, H.; Haentjens, P.; De Los Santos, M. J.; Fernandez-Sanchez, M.; Velasco, M.; Aebischer, P.; Devroey, P.; Simón, C. (2008).
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Glujovsky, Demián; Farquhar, Cindy; Quinteiro Retamar, Andrea Marta; Alvarez Sedo, Cristian Roberto; Blake, Deborah (2016-06-30). "Cleavage stage versus blastocyst stage embryo transfer in assisted reproductive technology".
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Mantikou, E.; Bontekoe, S.; Van Wely, M.; Seshadri, S.; Repping, S.; Mastenbroek, S. (2013). "Low oxygen concentrations for embryo culture in assisted reproductive technologies".
79:, and there is no evidence of a difference between the groups in cumulative pregnancy rates. Transfer day 2 instead of day 3 after fertilization has no differences in 137:(on top of a layer of cells from the woman's own uterine lining). With artificial culture medium, there can either be the same culture medium throughout the period ( 149:
A review in 2013 meta-analysis of commercially available IVF culture media was unable to identify a specific media that was superior in terms of pregnancy outcome.
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CO₂ / bicarbonate - buffered medium: uses the same physiological buffering system surrounding mammalian cells. Require the use of CO₂ incubators at 5-7%;
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Sunde, Arne; Brison, Daniel; Dumoulin, John; Harper, Joyce; Lundin, Kersti; Magli, M. Cristina; Van den Abbeel, Etienne; Veiga, Anna (October 2016).
613:"Advances in embryo culture platforms: novel approaches to improve preimplantation embryo development through modifications of the microenvironment" 205:
In addition to human embryo culture, the technique is employed extensively for non-human species, especially when exploring embryo development,
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Control and regulation of pH are mandatory for in vitro embryo culture. Culture media can be classified according to type of buffer used:
520: 281:"Neonatal outcomes among singleton births after blastocyst versus cleavage stage embryo transfer: a systematic review and meta-analysis" 744: 152:
Usage of low oxygen concentrations of 5% rather than about 20% in the atmosphere has been shown to increase live birth rate to a
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Embryos which reach the day 3 cell stage can be tested for chromosomal or specific genetic defects prior to possible transfer by
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Phosphate-buffered medium: does not require CO₂ environment. Seems to have detrimental effects in embryo development in vitro;
1048: 427:"Monozygotic twinning is not increased after single blastocyst transfer compared with single cleavage-stage embryo transfer" 217:
embryos, in particular, are frequently cultured for these specific research purposes. The two often used cultural media are
539:"Single or sequential medium are equally effective for the culture of human embryos to the blastocyst stage: a pilot study" 704:
Mantikou, E.; Youssef, M. A. F. M.; Van Wely, M.; Van Der Veen, F.; Al-Inany, H. G.; Repping, S.; Mastenbroek, S. (2013).
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MOPS-buffered medium: like HEPES, has the potential advantage that the buffering capacity is less temperature dependent.
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Wigger, Magdalena; Schneider, Marco; Feldmann, Anni; Assenmacher, Sonja; Zevnik, Branko; Tröder, Simon E. (2023-08-24).
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Melanie R. Freeman and Don Rieger. Nashville Fertility Center, Nashville, TN, U.S.A. and LifeGlobal, Guelph, ON, Canada
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of 1.24, without any evidence of increased risk for multiple pregnancies, miscarriages or congenital abnormalities.
206: 134: 1070: 370:"Day three versus day two embryo transfer following in vitro fertilization or intracytoplasmic sperm injection" 784:"New pH-buffering system for media utilized during gamete and embryo manipulations for assisted reproduction" 518:
Comparison Of A Single Medium With Sequential Media For Development Of Human Embryos To The Blastocyst Stage
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drawback that has to be taken into account, in fact, after day five embryo fusion is more likely to happen.
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abnormalities in embryos having undergone embryo culture, indicating a need to optimize the procedures.
572:"Embryo quality and implantation rate in two different culture media: ISM1 versus Universal IVF Medium" 228: 983:"Successful use of HTF as a basal fertilization medium during SEcuRe mouse in vitro fertilization" 112:
1.3) among births from embryos cultured until the blastocyst stage compared with cleavage stage.
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HEPES-buffered medium: used as buffered medium for human oocyte collection and embryo handling;
248: 20: 655:"An in vivo culture system for human embryos using an encapsulation technology: A pilot study" 517: 90: 255:
system, M2 medium facilitates embryo handling in ambient conditions without the need for CO
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where in resultant embryos are allowed to grow for some time in an artificial medium.
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Culture of embryos can either be performed in an artificial culture medium or in an
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The duration of embryo culture can be varied, conferring different stages of
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Baak, NA; Cantineau, AE; Farquhar, C; Brison, DR (17 September 2019).
706:"Embryo culture media and IVF/ICSI success rates: A systematic review" 569: 703: 425:
Papanikolaou EG, Fatemi H, Venetis C, et al. (February 2009).
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degrees. In addition, the pH levels should be between 7.2 and 7.5.
980: 926:"A brief history of the development of the KSOM family of media" 746: 570:
Xella S, Marsella T, Tagliasacchi D, et al. (April 2010).
115: 252: 48:. The main stages at which embryo transfer is performed are 830: 651: 833:"Temperature of embryo culture for assisted reproduction" 468: 424: 89:
is not increased after blastocyst transfer compared with
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Dar, S.; Lazer, T.; Shah, P. S.; Librach, C. L. (2014).
240: 471:"Time to take human embryo culture seriously: Table I" 200: 879: 536: 882:"Culture of oocytes and risk of imprinting defects" 278: 1041:Manipulating the mouse embryo: a laboratory manual 537:Schneider, D.T.; Verza, S.; Esteves, S.C. (2009). 464: 462: 368:Brown, Julie; Daya, Salim; Matson, Phill (2016). 247:incubator environment but is employed during the 1062: 243:. As with KSOM, HTF is only appropriate for a CO 367: 880:Anckaert, E.; De Rycke, M.; Smitz, J. (2012). 782:Swain, Jason E.; Pool, Thomas B. (June 2009). 459: 274: 272: 930:Journal of Assisted Reproduction and Genetics 116:Characteristics of an optimal embryo culture 837:The Cochrane Database of Systematic Reviews 374:The Cochrane Database of Systematic Reviews 331:The Cochrane Database of Systematic Reviews 269: 610: 1016: 998: 957: 897: 856: 799: 781: 721: 678: 628: 587: 554: 486: 442: 401: 296: 923: 96:There are significantly higher odds of 1063: 919: 917: 201:Embryo culture in non-human species 159: 13: 924:Summers, Michael C. (2013-09-18). 914: 219:potassium simplex optimized medium 14: 1082: 556:10.1016/j.fertnstert.2009.07.1564 69:preimplantation genetic diagnosis 589:10.1016/j.fertnstert.2008.12.030 444:10.1016/j.fertnstert.2008.12.088 231:mechanism, it is dependent on a 207:assisted reproductive technology 135:autologous endometrial coculture 1033: 974: 873: 824: 788:Reproductive Biomedicine Online 775: 738: 695: 645: 849:10.1002/14651858.CD012192.pub2 604: 563: 530: 511: 418: 386:10.1002/14651858.CD004378.pub3 361: 339:10.1002/14651858.CD002118.pub5 321: 179: 1: 801:10.1016/s1472-6483(10)60029-6 262: 193:Animal studies have detected 128: 211:genetically modified animals 7: 611:Swain JE, Smith GD (2011). 227:(HTF). Because KSOM uses a 29: 10: 1087: 1000:10.1186/s13104-023-06452-6 33: 942:10.1007/s10815-013-0097-8 886:Human Reproduction Update 749:Human Reproduction Update 710:Human Reproduction Update 285:Human Reproduction Update 209:, and the generation of 188: 60:stage (day 5 or 6 after 543:Fertility and Sterility 251:process. Buffered by a 1071:In vitro fertilisation 239:to maintain the right 899:10.1093/humupd/dms042 761:10.1093/humupd/dms055 723:10.1093/humupd/dms061 671:10.1093/humrep/dep005 630:10.1093/humupd/dmr006 488:10.1093/humrep/dew157 298:10.1093/humupd/dmu001 229:bicarbonate buffering 91:cleavage-stage embryo 34:Further information: 154:relative probability 106:congenital anomalies 87:Monozygotic twinning 617:Hum. Reprod. Update 987:BMC Research Notes 659:Human Reproduction 523:2012-03-21 at the 475:Human Reproduction 139:monoculture medium 52:(day 2 to 4 after 19:is a component of 1050:978-1-936113-00-2 481:(10): 2174–2182. 224:human tubal fluid 143:sequential system 1078: 1055: 1054: 1037: 1031: 1030: 1020: 1002: 978: 972: 971: 961: 921: 912: 911: 901: 877: 871: 870: 860: 828: 822: 821: 803: 779: 773: 772: 742: 736: 735: 725: 699: 693: 692: 682: 649: 643: 642: 632: 608: 602: 601: 591: 567: 561: 560: 558: 549:(3): S231–S232. 534: 528: 515: 509: 508: 490: 466: 457: 456: 446: 422: 416: 415: 405: 380:(12): CD004378. 365: 359: 358: 325: 319: 318: 300: 276: 160:Buffering system 1086: 1085: 1081: 1080: 1079: 1077: 1076: 1075: 1061: 1060: 1059: 1058: 1051: 1039: 1038: 1034: 979: 975: 922: 915: 878: 874: 843:(9): CD012192. 829: 825: 780: 776: 743: 739: 700: 696: 650: 646: 609: 605: 568: 564: 535: 531: 525:Wayback Machine 516: 512: 467: 460: 423: 419: 366: 362: 333:(6): CD002118. 326: 322: 277: 270: 265: 258: 246: 236: 203: 191: 182: 162: 131: 118: 81:live birth rate 77:embryo transfer 73:live birth rate 46:embryo transfer 38: 36:Embryo transfer 32: 12: 11: 5: 1084: 1074: 1073: 1057: 1056: 1049: 1032: 973: 936:(8): 995–999. 913: 872: 823: 794:(6): 799–810. 774: 737: 716:(3): 210–220. 694: 665:(4): 790–796. 644: 603: 582:(6): 1859–63. 576:Fertil. Steril 562: 529: 510: 458: 431:Fertil. Steril 417: 360: 320: 291:(3): 439–448. 267: 266: 264: 261: 256: 244: 234: 202: 199: 190: 187: 181: 178: 161: 158: 130: 127: 117: 114: 50:cleavage stage 31: 28: 17:Embryo culture 9: 6: 4: 3: 2: 1083: 1072: 1069: 1068: 1066: 1052: 1046: 1042: 1036: 1028: 1024: 1019: 1014: 1010: 1006: 1001: 996: 992: 988: 984: 977: 969: 965: 960: 955: 951: 947: 943: 939: 935: 931: 927: 920: 918: 909: 905: 900: 895: 891: 887: 883: 876: 868: 864: 859: 854: 850: 846: 842: 838: 834: 827: 819: 815: 811: 807: 802: 797: 793: 789: 785: 778: 770: 766: 762: 758: 754: 750: 745: 741: 733: 729: 724: 719: 715: 711: 707: 702: 698: 690: 686: 681: 676: 672: 668: 664: 660: 656: 648: 640: 636: 631: 626: 623:(4): 541–57. 622: 618: 614: 607: 599: 595: 590: 585: 581: 577: 573: 566: 557: 552: 548: 544: 540: 533: 526: 522: 519: 514: 506: 502: 498: 494: 489: 484: 480: 476: 472: 465: 463: 454: 450: 445: 440: 436: 432: 428: 421: 413: 409: 404: 399: 395: 391: 387: 383: 379: 375: 371: 364: 356: 352: 348: 344: 340: 336: 332: 324: 316: 312: 308: 304: 299: 294: 290: 286: 282: 275: 273: 268: 260: 254: 250: 249:fertilisation 242: 238: 230: 226: 225: 220: 216: 212: 208: 198: 196: 186: 177: 174: 171: 168: 165: 157: 155: 150: 147: 144: 140: 136: 126: 122: 113: 111: 107: 103: 99: 98:preterm birth 94: 92: 88: 84: 82: 78: 74: 70: 65: 63: 62:co-incubation 59: 55: 54:co-incubation 51: 47: 43: 42:embryogenesis 37: 27: 25: 24:fertilisation 23: 18: 1040: 1035: 990: 986: 976: 933: 929: 892:(1): 52–66. 889: 885: 875: 840: 836: 826: 791: 787: 777: 752: 748: 740: 713: 709: 697: 662: 658: 647: 620: 616: 606: 579: 575: 565: 546: 542: 532: 513: 478: 474: 437:(2): 592–7. 434: 430: 420: 377: 373: 363: 330: 323: 288: 284: 259:regulation. 222: 204: 192: 183: 175: 172: 169: 166: 163: 151: 148: 142: 138: 132: 123: 119: 95: 85: 66: 39: 21: 16: 15: 221:(KSOM) and 180:Temperature 993:(1): 184. 755:(3): 209. 263:References 195:epigenetic 129:Techniques 110:odds ratio 102:odds ratio 93:transfer. 58:blastocyst 1009:1756-0500 950:1058-0468 810:1472-6491 497:0268-1161 394:1469-493X 347:1469-493X 307:1355-4786 237:incubator 104:1.3) and 56:) or the 1065:Category 1027:37620881 1018:10463834 968:24046024 908:23054129 867:31529804 818:19490784 769:23377864 732:23385469 689:19273881 639:21454356 598:19152877 521:Archived 505:27554442 453:19243755 412:27976360 355:27357126 315:24480786 141:), or a 30:Duration 22:in vitro 959:3790120 858:6748832 680:2656929 403:6463848 185:rates. 1047:  1025:  1015:  1007:  966:  956:  948:  906:  865:  855:  816:  808:  767:  730:  687:  677:  637:  596:  503:  495:  451:  410:  400:  392:  353:  345:  313:  305:  253:HEPES 215:Mouse 189:Risks 1045:ISBN 1023:PMID 1005:ISSN 964:PMID 946:ISSN 904:PMID 863:PMID 814:PMID 806:ISSN 765:PMID 728:PMID 685:PMID 635:PMID 594:PMID 501:PMID 493:ISSN 449:PMID 408:PMID 390:ISSN 351:PMID 343:ISSN 311:PMID 303:ISSN 75:per 1013:PMC 995:doi 954:PMC 938:doi 894:doi 853:PMC 845:doi 796:doi 757:doi 718:doi 675:PMC 667:doi 625:doi 584:doi 551:doi 483:doi 439:doi 398:PMC 382:doi 335:doi 293:doi 64:). 44:at 1067:: 1021:. 1011:. 1003:. 991:16 989:. 985:. 962:. 952:. 944:. 934:30 932:. 928:. 916:^ 902:. 890:19 888:. 884:. 861:. 851:. 839:. 835:. 812:. 804:. 792:18 790:. 786:. 763:. 753:19 751:. 726:. 714:19 712:. 708:. 683:. 673:. 663:24 661:. 657:. 633:. 621:17 619:. 615:. 592:. 580:93 578:. 574:. 547:92 545:. 541:. 499:. 491:. 479:31 477:. 473:. 461:^ 447:. 435:93 433:. 429:. 406:. 396:. 388:. 378:12 376:. 372:. 349:. 341:. 309:. 301:. 289:20 287:. 283:. 271:^ 241:pH 233:CO 213:. 83:. 1053:. 1029:. 997:: 970:. 940:: 910:. 896:: 869:. 847:: 841:9 820:. 798:: 771:. 759:: 734:. 720:: 691:. 669:: 641:. 627:: 600:. 586:: 559:. 553:: 507:. 485:: 455:. 441:: 414:. 384:: 357:. 337:: 317:. 295:: 257:2 245:2 235:2 108:( 100:(

Index

in vitro fertilisation
Embryo transfer
embryogenesis
embryo transfer
cleavage stage
co-incubation
blastocyst
co-incubation
preimplantation genetic diagnosis
live birth rate
embryo transfer
live birth rate
Monozygotic twinning
cleavage-stage embryo
preterm birth
odds ratio
congenital anomalies
odds ratio
autologous endometrial coculture
relative probability
epigenetic
assisted reproductive technology
genetically modified animals
Mouse
potassium simplex optimized medium
human tubal fluid
bicarbonate buffering
CO2 incubator
pH
fertilisation

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