{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:53:17Z","timestamp":1791420797443,"version":"4.3.3"},"reference-count":31,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP19H04778"],"award-info":[{"award-number":["JP19H04778"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP20H03261"],"award-info":[{"award-number":["JP20H03261"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP21H05288"],"award-info":[{"award-number":["JP21H05288"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP26K020167"],"award-info":[{"award-number":["JP26K020167"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP23K05782"],"award-info":[{"award-number":["JP23K05782"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJSP2138"],"award-info":[{"award-number":["JPMJSP2138"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007449","name":"Takeda Science Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007449","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Development, Growth &amp; Differentiation"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Mouse embryos undergo drastic morphological changes during implantation, but analysis of this process has been challenging because it occurs within the uterus. Various in\u00a0vitro culture systems supporting development beyond implantation have been established since the 1970s. More recently, 3D\u2010geec, which supports embryos from the peri\u2010implantation to egg cylinder stages while preserving their three\u2010dimensional organization, was developed as a useful platform for studying this critical period. Combining 3D\u2010geec with genome editing in zygotes or experimental manipulation of embryos should further facilitate mechanistic studies of these stages. In this study, to enable such approaches, we established an in\u00a0vitro culture protocol that supports continuous development of mouse embryos from the zygote to the egg cylinder stage. Embryos cultured from the zygote to the late blastocyst stage in KSOM\n                    <jats:sup>AA<\/jats:sup>\n                    medium were not compatible with 3D\u2010geec. These embryos exhibited reduced cell numbers, delayed development, and reduced mTORC1 activity, which regulates cell growth and proliferation. The use of Advanced IVC medium supplemented with 10% fetal bovine serum (FBS) during the blastocyst stage substantially rescued these defects, and the resulting late blastocysts underwent further development into egg cylinders in 3D\u2010geec. Furthermore, genome\u2010edited embryos generated from zygotes developed into egg cylinders using this protocol and were successfully subjected to time\u2010lapse imaging during 3D\u2010geec. Thus, this culture protocol enables continuous in\u00a0vitro development from the zygote to the egg cylinder stage and provides a platform for studying gene function and cellular dynamics during peri\u2010implantation and early postimplantation development.\n                  <\/jats:p>","DOI":"10.1111\/dgd.70070","type":"journal-article","created":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:35:12Z","timestamp":1791419712000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Continuous In\u00a0Vitro Culture Protocol for Mouse Embryos From the Zygote to the Egg Cylinder Stage"],"prefix":"10.1111","volume":"68","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-3810-9879","authenticated-orcid":false,"given":"Rui","family":"Shibata","sequence":"first","affiliation":[{"name":"Laboratory for Embryogenesis, Graduate School of Frontier Biosciences The University of Osaka  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5455-4218","authenticated-orcid":false,"given":"Hiromi","family":"Shimojo","sequence":"additional","affiliation":[{"name":"Laboratory for Embryogenesis, Graduate School of Frontier Biosciences The University of Osaka  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8752-6846","authenticated-orcid":false,"given":"Hiroshi","family":"Sasaki","sequence":"additional","affiliation":[{"name":"Laboratory for Embryogenesis, Graduate School of Frontier Biosciences The University of Osaka  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,7]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2013.0538"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2014.186"},{"key":"e_1_2_10_4_1","volume-title":"Manipulating the Mouse Embryo: A Laboratory Manual","author":"Behringer R.","year":"2014"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/s1472\u20106483(10)62111\u20106"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2022113280"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature20578"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0092\u20108674(03)00392\u20101"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1231143"},{"key":"e_1_2_10_10_1","first-page":"277","article-title":"The Mechanism of Mouse Egg\u2010Cylinder Morphogenesis in Vitro","volume":"61","author":"Copp A. J.","year":"1981","journal-title":"Journal of Embryology and Experimental Morphology"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep11315"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ydbio.2016.07.017"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.203091"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/239200a0"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0012\u20101606(73)90145\u20100"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0012\u20101606(79)90217\u20103"},{"issue":"1","key":"e_1_2_10_17_1","first-page":"235","article-title":"Development in Vitro of Mouse Embryos From the Two\u2010Cell Egg Stage to the Early Somite Stage","volume":"31","author":"Hsu Y. 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