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After vitrification and warming, the COCs were subjected to IVM for 24 hours. Both fresh and vitrified COCs were analyzed for chromatin status, mitochondrial activity, reactive oxygen species levels, integrity of TZPs, DNA damage, and the expression of\n                      <jats:italic toggle=\"yes\">MPS1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">BUB1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">MAD1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">CX43<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">ZP3<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      The metaphase II (MII) rates of COCs vitrified with SPp (38%) were significantly higher than those vitrified with AFP3p (10%) (\n                      <jats:italic toggle=\"yes\">p<\/jats:italic>\n                      &lt; 0.05). The fluorescence intensity for CM-H\n                      <jats:sub>2<\/jats:sub>\n                      DCFDA (30 \u00b1 3.2) and nitrite\/nitrate levels (10.6 \u00b1 1.6) were higher in AFP3p COCs (\n                      <jats:italic toggle=\"yes\">p<\/jats:italic>\n                      &lt; 0.05). The transzonal projections (TZPs) of SPp COCs were intact and showed less DNA damage (25 \u00b1 1.15) compared with those of AFP3p (43 \u00b1 3.9) COCs (\n                      <jats:italic toggle=\"yes\">p<\/jats:italic>\n                      &lt; 0.05). The expression of the\n                      <jats:italic toggle=\"yes\">MPS1<\/jats:italic>\n                      (SPp 0.3\u2009\u00b1\u20090.4; AFP3p 0.07\u2009\u00b1\u20090.06) and\n                      <jats:italic toggle=\"yes\">BUB1<\/jats:italic>\n                      (SPp: 0.2\u2009\u00b1\u20090.4; AFP3p 0.005\u2009\u00b1\u20090.005) genes was higher in vitrified COCs compared with fresh control COCs (0.001\u2009\u00b1\u20090.0006; 0.001\u2009\u00b1\u20094.0) (\n                      <jats:italic toggle=\"yes\">p<\/jats:italic>\n                      &lt; 0.05). On the other hand, the\n                      <jats:italic toggle=\"yes\">MAD1<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">CX43<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">ZP3<\/jats:italic>\n                      genes were expressed only in fresh oocytes.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion:<\/jats:title>\n                    <jats:p>Under the conditions tested, SPp was the most suitable protocol for vitrifying bovine COCs, guaranteeing good MII rates, maintaining TZP integrity and reducing DNA damage.<\/jats:p>\n                  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