{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,31]],"date-time":"2025-08-31T10:16:40Z","timestamp":1756635400503,"version":"3.41.2"},"reference-count":24,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T00:00:00Z","timestamp":1627257600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:p>A time-lapse system (TLS) with a well-of-the-well (WOW) dish, which allows individual identification and the possibility of autocrine and paracrine signaling between group-cultured embryos, has been widely used in clinic. However, there is a need to re-think the inclusion principles of human embryos in WOW-based TLS, especially for grade IV (G4) embryos, which are considered to potentially have detrimental effects on surrounding embryos. Here, we carried out a single-center, large-cohort, retrospective study, comprising 303 patients undergoing IVF (148 cases) and ICSI (155 cases), with a total of 3282 embryos, to compare embryonic development until the blastocyst stage in the group culture system with or without G4 embryos. Further, LC-MS\/MS was used to analyze the G1-G4 embryo secretome to understand the influence of G4 embryos on the group culture microenvironment. We proved that polypronuclear (PPN) embryos positively contribute to the development of the neighboring embryos through secretion of ILIAP, ITI-H4, and keratin. Existence of more than one G4 embryo had a negative effect on the other embryos (p &amp;lt; 0.05). Moreover, G4 embryos were found to secrete KLKB1 and VTDB, which might harm the neighboring embryos. Thus, our study clarified that when embryos are subjected to group culture in WOW-based TLS, the PPN-derived embryos need not be removed, and it is important to ensure that no more than one G4 embryo is present to avoid negative effects on the neighboring embryos.<\/jats:p>","DOI":"10.3389\/fendo.2021.549216","type":"journal-article","created":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T11:50:00Z","timestamp":1627300200000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["The Inclusion Principles of Human Embryos in the WOW-Based Time-Lapse System: A Retrospective Cohort Study"],"prefix":"10.3389","volume":"12","author":[{"given":"Yuqiong","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xilin","family":"Qian","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanrong","family":"Kuai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yang","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,26]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.110.085522","article-title":"Time-Lapse Cinematography-Compatible Polystyrene-Based Microwell Culture System: A Novel Tool for Tracking the Development of Individual Bovine Embryos","volume":"83","author":"Sugimura","year":"2010","journal-title":"Biol Reprod"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1530\/REP-08-0370","article-title":"Effect of the Microenvironment and Embryo Density on Developmental Characteristics and Gene Expression Profile of Bovine Preimplantative Embryos Cultured In Vitro","volume":"137","author":"Hoelker","year":"2009","journal-title":"Reproduction"},{"key":"B3","doi-asserted-by":"publisher","first-page":"CD011320","DOI":"10.1002\/14651858.CD011320.pub3","article-title":"Time-Lapse Systems for Embryo Incubation and Assessment in Assisted Reproduction","volume":"5","author":"Armstrong","year":"2018","journal-title":"Cochrane Database Syst Rev"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-018-1252-z","article-title":"A Microwell Culture System That Allows Group Culture and Is Compatible With Human Single Media","volume":"35","author":"Ieda","year":"2018","journal-title":"J Assist Reprod Genet"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1007\/s00404-017-4403-z","article-title":"Embryo Density may Affect Embryo Quality During In Vitro Culture in a Microwell Group Culture Dish","volume":"296","author":"Lehner","year":"2017","journal-title":"Arch Gynecol Obstet"},{"key":"B6","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1016\/S1472-6483(10)60296-9","article-title":"The Well-of-the-Well System: An Efficient Approach to Improve Embryo Development","volume":"17","author":"Vajta","year":"2008","journal-title":"Reprod BioMed Online"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1017\/S096719941400001X","article-title":"The Efficacy of the Well of the Well (WOW) Culture System on Development of Bovine Embryos in a Small Group and the Effect of Number of Adjacent Embryos on Their Development","volume":"23","author":"Kang","year":"2015","journal-title":"Zygote"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(200003)55:3<256::AID-MRD3>3.0.CO;2-7","article-title":"New Method for Culture of Zona-Included or Zona-Free Embryos: The Well of the Well (WOW) System","volume":"55","author":"Vajta","year":"2000","journal-title":"Mol Reprod Dev"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-012-9892-x","article-title":"Influence of Group Embryo Culture Strategies on the Blastocyst Development and Pregnancy Outcome","volume":"30","author":"Tao","year":"2013","journal-title":"J Assist Reprod Genet"},{"key":"B10","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.ydbio.2005.05.001","article-title":"Development of Porcine Embryos In Vivo and In Vitro; 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