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From ZHBTc4 ESCs, EBs were created by the hanging-drop method, starting with 1000 cells/drop. After 2 days in the drop state, each drop begins to grow on a non-coated sterile cover glass for immunocytochemistry, and on a round-shaped 96-well plate for live-cell imaging. In all culture situations, media were fed through addition per day and plate movement was also fixed to a minimum for minimizing the physical impact on the EBs. All EBs were cultured using ESC media without LIF.
Copyright and License information: ©2020 The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact moc.puo@snoissimrep.slanruoj
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