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Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System

To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in...

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Autores principales: SUGIMURA, Satoshi, AKAI, Tomonori, HASHIYADA, Yutaka, AIKAWA, Yoshio, OHTAKE, Masaki, MATSUDA, Hideo, KOBAYASHI, Shuji, KOBAYASHI, Eiji, KONISHI, Kazuyuki, IMAI, Kei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Society for Reproduction and Development 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934200/
https://www.ncbi.nlm.nih.gov/pubmed/23154384
http://dx.doi.org/10.1262/jrd.2012-113
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author SUGIMURA, Satoshi
AKAI, Tomonori
HASHIYADA, Yutaka
AIKAWA, Yoshio
OHTAKE, Masaki
MATSUDA, Hideo
KOBAYASHI, Shuji
KOBAYASHI, Eiji
KONISHI, Kazuyuki
IMAI, Kei
author_facet SUGIMURA, Satoshi
AKAI, Tomonori
HASHIYADA, Yutaka
AIKAWA, Yoshio
OHTAKE, Masaki
MATSUDA, Hideo
KOBAYASHI, Shuji
KOBAYASHI, Eiji
KONISHI, Kazuyuki
IMAI, Kei
author_sort SUGIMURA, Satoshi
collection PubMed
description To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 µl medium for 168 h. The blastocysts at Day 7 were analyzed for number of cells and expression of ten genes (CDX2, IFN-tau, PLAC8, NANOG, OCT4, SOX2, AKR1B1, ATP5A1, GLUT1 and IGF2R). In droplet culture, the rates of formation of >4-cell cleavage embryos and blastocysts were significantly lower in embryos cultured at 5 embryos per droplet than in those cultured at 15 or 25 embryos per droplet, but not in WOW culture. In both droplet and WOW culture, developmental kinetics and blastocyst cell numbers did not differ among any groups. IFN-tau expression in embryos cultured at 25 embryos per droplet was significantly higher than in those cultured at 15 embryos per droplet and in artificial insemination (AI)-derived blastocysts. Moreover, IGF2R expression was significantly lower in the 25-embryo group than in the 5-embryo group and in AI-derived blastocysts. In WOW culture, these expressions were not affected by embryo density and were similar to those in AI-derived blastocysts. These results suggest that, as compared with conventional droplet culture, in vitro development and expression of IFN-tau and IGF2R in the microwell system may be insensitive to embryo density.
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spelling pubmed-39342002014-03-06 Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System SUGIMURA, Satoshi AKAI, Tomonori HASHIYADA, Yutaka AIKAWA, Yoshio OHTAKE, Masaki MATSUDA, Hideo KOBAYASHI, Shuji KOBAYASHI, Eiji KONISHI, Kazuyuki IMAI, Kei J Reprod Dev Original Article To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 µl medium for 168 h. The blastocysts at Day 7 were analyzed for number of cells and expression of ten genes (CDX2, IFN-tau, PLAC8, NANOG, OCT4, SOX2, AKR1B1, ATP5A1, GLUT1 and IGF2R). In droplet culture, the rates of formation of >4-cell cleavage embryos and blastocysts were significantly lower in embryos cultured at 5 embryos per droplet than in those cultured at 15 or 25 embryos per droplet, but not in WOW culture. In both droplet and WOW culture, developmental kinetics and blastocyst cell numbers did not differ among any groups. IFN-tau expression in embryos cultured at 25 embryos per droplet was significantly higher than in those cultured at 15 embryos per droplet and in artificial insemination (AI)-derived blastocysts. Moreover, IGF2R expression was significantly lower in the 25-embryo group than in the 5-embryo group and in AI-derived blastocysts. In WOW culture, these expressions were not affected by embryo density and were similar to those in AI-derived blastocysts. These results suggest that, as compared with conventional droplet culture, in vitro development and expression of IFN-tau and IGF2R in the microwell system may be insensitive to embryo density. The Society for Reproduction and Development 2012-11-15 2013-04 /pmc/articles/PMC3934200/ /pubmed/23154384 http://dx.doi.org/10.1262/jrd.2012-113 Text en ©2013 Society for Reproduction and Development http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.
spellingShingle Original Article
SUGIMURA, Satoshi
AKAI, Tomonori
HASHIYADA, Yutaka
AIKAWA, Yoshio
OHTAKE, Masaki
MATSUDA, Hideo
KOBAYASHI, Shuji
KOBAYASHI, Eiji
KONISHI, Kazuyuki
IMAI, Kei
Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System
title Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System
title_full Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System
title_fullStr Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System
title_full_unstemmed Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System
title_short Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System
title_sort effect of embryo density on in vitro development and gene expression in bovine in vitro-fertilized embryos cultured in a microwell system
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934200/
https://www.ncbi.nlm.nih.gov/pubmed/23154384
http://dx.doi.org/10.1262/jrd.2012-113
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