title: Time-lapse cinematography-compatible polystyrene-based microwell culture system: a novel tool for tracking the development of individual bovine embryos.
abstract: We have developed a polystyrene-based well-of-the-well (WOW) system using injection molding to track individual embryos throughout culture using time-lapse cinematography (TLC). WOW culture of bovine embryos following in vitro fertilization was compared with conventional droplet culture (control). No differences between control- and WOW-cultured embryos were observed during development to the blastocyst stage. Morphological quality and inner cell mass (ICM) and trophectoderm (TE) cell numbers were not different between control- and WOW-derived blastocysts; however, apoptosis in both the ICM and TE cells was reduced in WOW culture (P < 0.01). Oxygen consumption in WOW-derived blastocysts was closer to physiological level than that of control-derived blastocysts. Moreover, WOW culture improved embryo viability, as indicated by increased pregnancy rates at Days 30 and 60 after embryo transfer (P < 0.05). TLC monitoring was performed to evaluate the cleavage pattern and the duration of the first cell cycle of embryos from oocytes collected by ovum pickup; correlations with success of pregnancy were determined. Logistic regression analysis indicated that the cleavage pattern correlated with success of pregnancy (P < 0.05), but cell cycle length did not. Higher pregnancy rates (66.7%) were observed for animals in which transferred blastocysts had undergone normal cleavage, identified by the presence of two blastomeres of the same size without fragmentation, than among those with abnormal cleavage (33.3%). These results suggest that our microwell culture system is a powerful tool for producing and selecting healthy embryos and for identifying viability biomarkers.
authors.affiliation: National Livestock Breeding Center, Nishigo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Sugimura
authors.forename: Tomonori
authors.initials: T
authors.lastname: Akai
authors.forename: Tamás
authors.initials: T
authors.lastname: Somfai
authors.forename: Muneyuki
authors.initials: M
authors.lastname: Hirayama
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Aikawa
authors.forename: Masaki
authors.initials: M
authors.lastname: Ohtake
authors.forename: Hideshi
authors.initials: H
authors.lastname: Hattori
authors.forename: Shuji
authors.initials: S
authors.lastname: Kobayashi
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hashiyada
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Konishi
authors.forename: Kei
authors.initials: K
authors.lastname: Imai
mesh_terms: D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D053624:Blastocyst Inner Cell Mass / Q000166:cytology; D002417:Cattle; D002452:Cell Count; D002453:Cell Cycle; D046149:Embryo Culture Techniques / Q000295:instrumentation* / Q000379:methods; D010064:Embryo Implantation; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro; D007700:Kinetics; D018715:Microscopy, Video; D010101:Oxygen Consumption; D011137:Polystyrenes; D011247:Pregnancy; D011256:Pregnancy Outcome; D059008:Time-Lapse Imaging; D014327:Trophoblasts / Q000166:cytology
chemical_list: D011137:Polystyrenes