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. 2021 Jan 8;11:61. doi: 10.1038/s41598-020-79732-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Lobactis scutaria reproduction. (a) When fresh sperm and fresh sperm + 10% DMSO were added to fresh eggs, the reproductive success was > 75% across all fresh sperm treatments. However, during cryopreservation the motile concentration was reduced by a factor of 10, thereby reducing the egg:sperm ratio to 1:5400 and 1:13,000 of motile sperm for the successful cryo trials. (b) Understanding the motile concentration of cryopreserved sperm and obtaining accurate egg:sperm ratios are critical for predicting robust fertilisation success with thawed sperm samples. This figure shows correlation between motile sperm concentration per egg (x-axis, logarithmic scale) and fertilisation success (y-axis) in eggs fertilised by fresh sperm in FSW, fresh sperm in 10% DMSO, and cryopreserved sperm in 10% DMSO. The correlation coefficient r between the two variables is 0.75 (P = 8.6 × 10⁻¹⁰). Fresh sperm samples from 6 males were pooled and mixed with individual females (n = 6). One and two data points are missing in the fresh sperm + 10% DMSO, 100 and 200 µL categories, respectively. Bars in panel a are SE.