Ex utero mouse embryogenesis from pre-gastrulation to late organogenesis

Alejandro Aguilera-Castrejon (), Bernardo Oldak, Tom Shani, Nadir Ghanem, Chen Itzkovich, Sharon Slomovich, Shadi Tarazi, Jonathan Bayerl, Valeriya Chugaeva, Muneef Ayyash, Shahd Ashouokhi, Daoud Sheban, Nir Livnat, Lior Lasman, Sergey Viukov, Mirie Zerbib, Yoseph Addadi, Yoach Rais, Saifeng Cheng, Yonatan Stelzer, Hadas Keren-Shaul, Raanan Shlomo, Rada Massarwa, Noa Novershtern, Itay Maza () and Jacob H. Hanna ()
Additional contact information
Alejandro Aguilera-Castrejon: Weizmann Institute of Science
Bernardo Oldak: Weizmann Institute of Science
Tom Shani: Weizmann Institute of Science
Nadir Ghanem: Rambam Health Care Campus
Chen Itzkovich: Rambam Health Care Campus
Sharon Slomovich: Israel Institute of Technology - Technion
Shadi Tarazi: Weizmann Institute of Science
Jonathan Bayerl: Weizmann Institute of Science
Valeriya Chugaeva: Weizmann Institute of Science
Muneef Ayyash: Weizmann Institute of Science
Shahd Ashouokhi: Weizmann Institute of Science
Daoud Sheban: Weizmann Institute of Science
Nir Livnat: Weizmann Institute of Science
Lior Lasman: Weizmann Institute of Science
Sergey Viukov: Weizmann Institute of Science
Mirie Zerbib: Weizmann Institute of Science
Yoseph Addadi: Weizmann Institute of Science
Yoach Rais: Weizmann Institute of Science
Saifeng Cheng: Weizmann Institute of Science
Yonatan Stelzer: Weizmann Institute of Science
Hadas Keren-Shaul: Weizmann Institute of Science
Raanan Shlomo: Arad Technologies Ltd
Rada Massarwa: Weizmann Institute of Science
Noa Novershtern: Weizmann Institute of Science
Itay Maza: Israel Institute of Technology - Technion
Jacob H. Hanna: Weizmann Institute of Science

Nature, 2021, vol. 593, issue 7857, 119-124

Abstract: Abstract The mammalian body plan is established shortly after the embryo implants into the maternal uterus, and our understanding of post-implantation developmental processes remains limited. Although pre- and peri-implantation mouse embryos are routinely cultured in vitro1,2, approaches for the robust culture of post-implantation embryos from egg cylinder stages until advanced organogenesis remain to be established. Here we present highly effective platforms for the ex utero culture of post-implantation mouse embryos, which enable the appropriate development of embryos from before gastrulation (embryonic day (E) 5.5) until the hindlimb formation stage (E11). Late gastrulating embryos (E7.5) are grown in three-dimensional rotating bottles, whereas extended culture from pre-gastrulation stages (E5.5 or E6.5) requires a combination of static and rotating bottle culture platforms. Histological, molecular and single-cell RNA sequencing analyses confirm that the ex utero cultured embryos recapitulate in utero development precisely. This culture system is amenable to the introduction of a variety of embryonic perturbations and micro-manipulations, the results of which can be followed ex utero for up to six days. The establishment of a system for robustly growing normal mouse embryos ex utero from pre-gastrulation to advanced organogenesis represents a valuable tool for investigating embryogenesis, as it eliminates the uterine barrier and allows researchers to mechanistically interrogate post-implantation morphogenesis and artificial embryogenesis in mammals.

Date: 2021
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DOI: 10.1038/s41586-021-03416-3

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