Shoko Ieda1, Tomonori Akai2, Yoko Sakaguchi1, Sumi Shimamura1, Atsushi Sugawara3,4, Masahiro Kaneda5, Satoko Matoba6, Masanori Kagota2, Satoshi Sugimura7, Hirotsune Kaijima8. 1. Minatomirai Yume Clinic, Yokohama, 3-6-3 MM Park Building 2nd Floor Minatomirai, Nishi-ku, Yokohama, Kanagawa, 220-0012, Japan. 2. Dai Nippon Printing Co., Ltd., Chiba, 277-0871, Japan. 3. Department of Biological Production, Tokyo University of Agriculture and Technology, Tokyo, 183-8509, Japan. 4. John A. Burns School of Medicine, Department of Anatomy, Institute for Biogenesis Research, University of Hawaii, Honolulu, HI, 96822, USA. 5. Division of Animal Life Science, Tokyo University of Agriculture and Technology, Tokyo, 183-8509, Japan. 6. Animal Breeding and Reproduction Research Division, National Agriculture and Food Research Organization (NARO) Institute of Livestock and Grassland Science, Ibaraki, 305-0901, Japan. 7. Department of Biological Production, Tokyo University of Agriculture and Technology, Tokyo, 183-8509, Japan. satoshis@cc.tuat.ac.jp. 8. Minatomirai Yume Clinic, Yokohama, 3-6-3 MM Park Building 2nd Floor Minatomirai, Nishi-ku, Yokohama, Kanagawa, 220-0012, Japan. h-kaijima@mm-yumeclinic.com.
Abstract
PURPOSE: A microwell culture system that facilitates group culture, such as well-of-the-well (WOW), improves embryonic development in an individual culture. We examined the effect of WOW on embryonic development in vitro with commercially available human single culture media. METHODS: Using four different commercial human single culture media, in vitro development and imprinted gene expression of bovine embryos cultured in WOW were compared to droplet culture (one zygote per drop). To determine the effects of microwell and group culture on embryonic development, different numbers of embryos were cultured in droplet or WOW. Diffusion simulation of accumulating metabolites was conducted using the finite volume method. RESULTS: WOW had a positive effect on bovine embryonic development, regardless of the type of single culture media. Imprinted gene expression was not different between droplet- and WOW-derived blastocysts. The microwell and group cultures in WOW showed a significant positive effect on the rate of total blastocysts and the rate of development to the expanded and hatching blastocyst stages. The assumed cumulative metabolite concentration of WOW with one embryo was 1.47 times higher than that of droplet culture with one embryo. Furthermore, the concentration of WOW with three embryos was 1.54 times higher than that of WOW with one embryo. CONCLUSIONS: In using human single culture media, a microwell culture system that allows group culture could be a powerful clinical tool for improving the success of assisted reproductive technologies.
PURPOSE: A microwell culture system that facilitates group culture, such as well-of-the-well (WOW), improves embryonic development in an individual culture. We examined the effect of WOW on embryonic development in vitro with commercially available human single culture media. METHODS: Using four different commercial human single culture media, in vitro development and imprinted gene expression of bovine embryos cultured in WOW were compared to droplet culture (one zygote per drop). To determine the effects of microwell and group culture on embryonic development, different numbers of embryos were cultured in droplet or WOW. Diffusion simulation of accumulating metabolites was conducted using the finite volume method. RESULTS: WOW had a positive effect on bovine embryonic development, regardless of the type of single culture media. Imprinted gene expression was not different between droplet- and WOW-derived blastocysts. The microwell and group cultures in WOW showed a significant positive effect on the rate of total blastocysts and the rate of development to the expanded and hatching blastocyst stages. The assumed cumulative metabolite concentration of WOW with one embryo was 1.47 times higher than that of droplet culture with one embryo. Furthermore, the concentration of WOW with three embryos was 1.54 times higher than that of WOW with one embryo. CONCLUSIONS: In using human single culture media, a microwell culture system that allows group culture could be a powerful clinical tool for improving the success of assisted reproductive technologies.
Entities:
Keywords:
Bovine embryo assay; Group culture; Human single medium; Metabolites; Microwell
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