Department of Animal Science, Sangji University, Wonju, 26339, Republic of Korea.
Department of Companion Animal, Catholic Sangji College, Andong, 36686, Republic of Korea.
Vet Res Commun. 2023 Dec;47(4):2101-2109. doi: 10.1007/s11259-023-10160-8. Epub 2023 Jul 11.
Immunological approaches are gaining attention as a convenient and economical method for sex-sorting mammalian spermatozoa. A monoclonal antibody (WholeMom™) has previously been reported to cause agglutination of Y-chromosome-bearing spermatozoa in frozen-thawed semen for gender preselection. However, its usefulness for gender preselection in fresh semen and subsequent in vitro fertilization (IVF) after freeze-thawing has not been reported. This study investigated the in vitro development of cattle embryos produced from fresh bull semen pre-treated with WholeMom™ monoclonal antibody. Results showed that antibody-treated, non-agglutinated spermatozoa (presumably X-chromosome-bearing spermatozoa) could fertilize cattle oocytes in vitro. However, embryos generated from non-agglutinated (enriched in X-chromosome-bearing spermatozoa) had a lower (p < 0.05) ability to cleave (66.4 ± 2.5% vs. 75.1 ± 3.3%) than those of non-treated control sperm. Nevertheless, the percentage of blastocysts developed from cleaved embryos did not differ (p > 0.05) between the groups (34.8 ± 3.7% vs. 35.8 ± 3.4%). Duplex PCR of blastocysts, using a bovine-specific universal primer pair and a Y-chromosome-specific primer pair, showed a sex ratio of 95.8% females from sex-sorted spermatozoa, which was higher than those of non-treated control spermatozoa (46.4%). In conclusion, the results of the present study suggest that monoclonal antibody-based enrichment of X- chromosome-bearing spermatozoa can be applied to fresh bull semen without compromising their post-fertilization early embryonic development to the blastocyst stage. Future studies should investigate the term development and sex ratio of calves from antibody-treated spermatozoa.
免疫方法作为一种方便、经济的哺乳动物精子性别分选方法受到关注。先前有报道称，一种单克隆抗体（WholeMom™）可引起冷冻解冻精液中携带 Y 染色体的精子凝集，从而进行性别预选。然而，其在新鲜精液中的性别预选和随后的冷冻解冻后体外受精（IVF）中的用途尚未报道。本研究调查了用 WholeMom™单克隆抗体预处理新鲜公牛精液后产生的牛胚胎的体外发育情况。结果表明，抗体处理后未凝集的精子（推测为携带 X 染色体的精子）可以体外受精牛卵母细胞。然而，来自未凝集的（富含携带 X 染色体的精子）胚胎的分裂能力较低（p<0.05）（66.4±2.5%比 75.1±3.3%），低于未处理对照组的精子。然而，从分裂胚胎中发育的囊胚的百分比在两组之间没有差异（p>0.05）（34.8±3.7%比 35.8±3.4%）。使用牛特异性通用引物对和 Y 染色体特异性引物对的囊胚双重 PCR 分析显示，从分选的精子中获得的雌性胚胎性别比为 95.8%，高于未处理对照组的精子（46.4%）。总之，本研究结果表明，基于单克隆抗体的 X 染色体携带精子富集可应用于新鲜公牛精液，而不会影响其受精后早期胚胎发育至囊胚阶段。未来的研究应调查抗体处理精子的小牛的期末发育和性别比。
