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

Autor:	M. Ohtake, Kazuyuki Konishi, Tomonori Akai, Yutaka Hashiyada, Hideo Matsuda, Shuji Kobayashi, Y. Aikawa, Satoshi Sugimura, Kei Imai, Eiji Kobayashi
Rok vydání:	2013
Předmět:	Regulation of gene expression Homeobox protein NANOG animal structures Embryogenesis Embryo Biology Cleavage (embryo) Molecular biology In vitro Andrology medicine.anatomical_structure embryonic structures Gene expression medicine Animal Science and Zoology Blastocyst
Zdroj:	Journal of Reproduction and Development. 59:115-122
ISSN:	1348-4400 0916-8818
DOI:	10.1262/jrd.2012-113
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::46c98ab30baf3ef9323231a4f7ba38ed https://doi.org/10.1262/jrd.2012-113 Zobrazit plný text záznamu