To explore the blastocyst development potency of three-pronuclear (PN) zygotes after one unnecessary pronucleus reduction by micropuncture. Methods: Between July and October 2018, 151 infertile women with 3PN zygotes who had experinced short-term in vitro fertilization (IVF) after one unnecessary pronucleus reduction by micropuncture were included in experimental group, and 414 women with normal 2PN zygotes were in control group. The embryonic development situation of the women was compared between the two groups. The available cells of blastaea trophectoderm (TE) and inner cell mass (ICM) of the women in the experimental group were collected for high-throughput sequencing and copy number variation (CNV) analysis. In the experimental group, 141 of 151 3PNs zygotes of the women were micronucleated successfully, with 93.4% success rate of enucleation. The cleavage rate on the 3rd day (77.3%) and the blastocyst formation rate on the 3rd to 5th (97.3%) of the women in the experimental group were significant lower than those (8.3% and 62.6%) of the women in the control group (P<0.05). The rate of embryogenesis with ≥6 cells (87.2%) and the high quality embryos rate (78.4%) of the women in the experimental group were significant higher than those (80.7% and 56.8%) of the women in the control group (P<0.05). The available blastocyst rate (88.9%) of the women in the experimental group was similar to that (63.2%) of the women in the control group (P>0.05). A total of 6 TE samples and 5 ICM samples were detected, and among them, karyotype test results of 10 samples were 46XN, and 1 TE sample was 46,XN,-6(×1, MOS, -40%), -6q  (×1, MOS,-50%). Conclusion: After correcting one unnecessary pronucleu of 3PN zygotes by micropuncture of IVF women, their embryo developmental potential have been improved, but its utilization value needs further study.