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However, concerns persist regarding the safety and efficacy of AOA in clinical practice. We demonstrated that single\u2010pulse nanosecond pulsed electric field (nsPEF) stimulation induced Ca\n                    <jats:sup>2+<\/jats:sup>\n                    signaling patterns that depend on intensity in both mouse and human oocytes, facilitating parthenogenetic activation and blastocyst formation. The sperm\u2010initiated physiological Ca\n                    <jats:sup>2+<\/jats:sup>\n                    oscillations were effectively replicated by a series of Ca\n                    <jats:sup>2+<\/jats:sup>\n                    signals triggered by repeated nsPEF at low or medium intensities, resulting in a significantly higher developmental potential for activated oocytes compared to those treated with A23187 (78.13% vs. 26.70%). The nsPEF stimulation achieved precise manipulation of calcium signaling through two distinct mechanisms: low\u2010intensity nsPEF pulses mediated repetitive extracellular Ca\n                    <jats:sup>2+<\/jats:sup>\n                    influx in an electro\u2010permeable manner, while medium\u2010intensity nsPEF stimulation triggered periodic Ca\n                    <jats:sup>2+<\/jats:sup>\n                    release from the endoplasmic reticulum via the PIP\n                    <jats:sub>2<\/jats:sub>\n                    \u2013IP\n                    <jats:sub>3<\/jats:sub>\n                    \u2013IP\n                    <jats:sub>3<\/jats:sub>\n                    R pathway, generating intracellular Ca\n                    <jats:sup>2+<\/jats:sup>\n                    oscillations that resemble physiological patterns. The non\u2010invasive nsPEF procedure ensured the safety of oocyte activation by maintaining cellular integrity and minimizing stress responses. The efficacy of nsPEF exposure in precisely manipulating Ca\n                    <jats:sup>2+<\/jats:sup>\n                    signaling patterns is also demonstrated in human mature oocytes. This study establishes a quantitative, non\u2010invasive nsPEF protocol for AOA that mimics the activation signaling delivered by sperm. This innovative approach overcomes the limitations of conventional chemical activators by enhancing biosafety and clinical efficacy, particularly for patients experiencing total fertilization failure due to severe male infertility. Its ability to accurately regulate Ca\n                    <jats:sup>2+<\/jats:sup>\n                    signaling presents significant potential for advancing research in various fields, including embryonic development and germ cell differentiation.\n                  <\/jats:p>","DOI":"10.1002\/btm2.70094","type":"journal-article","created":{"date-parts":[[2025,12,6]],"date-time":"2025-12-06T04:30:10Z","timestamp":1764995410000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Precise mimicry of physiological Ca\n                    <sup>2+<\/sup>\n                    oscillations for mammalian oocyte activation by nanosecond pulsed electric field"],"prefix":"10.1002","volume":"11","author":[{"given":"Yi\u2010Dan","family":"Sun","sequence":"first","affiliation":[{"name":"State Key Laboratory of Animal Biotech Breeding, College of Biological Sciences China Agricultural University  Beijing 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