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25 August 2010 Time-Lapse Cinematography-Compatible Polystyrene-Based Microwell Culture System: A Novel Tool for Tracking the Development of Individual Bovine Embryos
Satoshi Sugimura, Tomonori Akai, Tamás Somfai, Muneyuki Hirayama, Yoshio Aikawa, Masaki Ohtake, Hideshi Hattori, Shuji Kobayashi, Yutaka Hashiyada, Kazuyuki Konishi, Kei Imai
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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.

Satoshi Sugimura, Tomonori Akai, Tamás Somfai, Muneyuki Hirayama, Yoshio Aikawa, Masaki Ohtake, Hideshi Hattori, Shuji Kobayashi, Yutaka Hashiyada, Kazuyuki Konishi, and Kei Imai "Time-Lapse Cinematography-Compatible Polystyrene-Based Microwell Culture System: A Novel Tool for Tracking the Development of Individual Bovine Embryos," Biology of Reproduction 83(6), 970-978, (25 August 2010). https://doi.org/10.1095/biolreprod.110.085522
Received: 22 April 2010; Accepted: 1 August 2010; Published: 25 August 2010
KEYWORDS
bovine embryos
cleavage pattern
developmental biology
embryo
in vitro fertilization
ovum pickup/transport
pregnancy
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