Ex Utero Culture of Mouse Embryos from Pregastrulation to Advanced Organogenesis. 2021

Alejandro Aguilera-Castrejon, and Jacob H Hanna
Department of Molecular Genetics, Weizmann Institute of Science; alejandroac@weizmann.ac.il.

Postimplantation mammalian embryo culture methods have been generally inefficient and limited to brief periods after dissection out of the uterus. Platforms have been recently developed for highly robust and prolonged ex utero culture of mouse embryos from egg-cylinder stages until advanced organogenesis. These platforms enable appropriate and faithful development of pregastrulating embryos (E5.5) until the hind limb formation stage (E11). Late gastrulating embryos (E7.5) are grown in rotating bottles in these settings, while extended culture from pregastrulation stages (E5.5 or E6.5) requires a combination of static and rotating bottle cultures. In addition, sensitive regulation of O2 and CO2 concentration, gas pressure, glucose levels, and the use of a specific ex utero culture medium are critical for proper embryo development. Here, a detailed step-by-step protocol for extended ex utero mouse embryo culture is provided. The ability to grow normal mouse embryos ex utero from gastrulation to organogenesis represents a valuable tool for characterizing the effect of different experimental perturbations during embryonic development.

UI MeSH Term Description Entries
D008322 Mammals Warm-blooded vertebrate animals belonging to the class Mammalia, including all that possess hair and suckle their young. Mammalia,Mammal
D011247 Pregnancy The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH. Gestation,Pregnancies
D004622 Embryo, Mammalian The entity of a developing mammal (MAMMALS), generally from the cleavage of a ZYGOTE to the end of embryonic differentiation of basic structures. For the human embryo, this represents the first two months of intrauterine development preceding the stages of the FETUS. Embryonic Structures, Mammalian,Mammalian Embryo,Mammalian Embryo Structures,Mammalian Embryonic Structures,Embryo Structure, Mammalian,Embryo Structures, Mammalian,Embryonic Structure, Mammalian,Embryos, Mammalian,Mammalian Embryo Structure,Mammalian Embryonic Structure,Mammalian Embryos,Structure, Mammalian Embryo,Structure, Mammalian Embryonic,Structures, Mammalian Embryo,Structures, Mammalian Embryonic
D005260 Female Females
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D046149 Embryo Culture Techniques The technique of maintaining or growing mammalian EMBRYOS in vitro. This method offers an opportunity to observe EMBRYONIC DEVELOPMENT; METABOLISM; and susceptibility to TERATOGENS. Blastocyst Culture Techniques,Blastocyst Culture Technique,Culture Technique, Blastocyst,Culture Technique, Embryo,Culture Techniques, Blastocyst,Culture Techniques, Embryo,Embryo Culture Technique
D047108 Embryonic Development Morphological and physiological development of EMBRYOS. Embryo Development,Embryogenesis,Postimplantation Embryo Development,Preimplantation Embryo Development,Embryonic Programming,Post-implantation Embryo Development,Postnidation Embryo Development,Postnidation Embryo Development, Animal,Pre-implantation Embryo Development,Prenidation Embryo Development, Animal,Development, Embryo,Development, Embryonic,Development, Postnidation Embryo,Embryo Development, Post-implantation,Embryo Development, Postimplantation,Embryo Development, Postnidation,Embryo Development, Pre-implantation,Embryo Development, Preimplantation,Embryonic Developments,Embryonic Programmings,Post implantation Embryo Development,Pre implantation Embryo Development
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D054262 Gastrulation A process of complicated morphogenetic cell movements that reorganizes a bilayer embryo into one with three GERM LAYERS and specific orientation (dorsal/ventral; anterior/posterior). Gastrulation describes the germ layer development of a non-mammalian BLASTULA or that of a mammalian BLASTOCYST.
D038081 Organogenesis Formation of differentiated cells and complicated tissue organization to provide specialized functions.

Related Publications

Alejandro Aguilera-Castrejon, and Jacob H Hanna
January 2019, Methods in molecular biology (Clifton, N.J.),
Alejandro Aguilera-Castrejon, and Jacob H Hanna
January 2011, Methods in molecular biology (Clifton, N.J.),
Alejandro Aguilera-Castrejon, and Jacob H Hanna
October 2022, Cellular reprogramming,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
May 2023, Cell,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
January 1979, Journal of embryology and experimental morphology,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
September 2022, Nature methods,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
November 2022, Artificial organs,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
July 2023, Nature methods,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
November 2010, Cell,
Alejandro Aguilera-Castrejon, and Jacob H Hanna
March 2015, Journal of visualized experiments : JoVE,
Copied contents to your clipboard!