期刊
ZYGOTE
卷 -, 期 -, 页码 -出版社
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S0967199423000096
关键词
Artificial oocytes; Meiosis; Somatic cell haploidization; Somatic cell nuclear transfer; Time-lapse microscopy
Oocyte-mediated somatic cell haploidization is a feasible technique for creating functional gametes in mammals. Experimental data have shown successful reconstruction of murine oocytes, resulting in a blastocyst formation rate of 12.7% and a diploid embryo rate of 27.3%. This method has the potential to generate patient-genotyped oocytes, but further research is needed to improve the generation of chromosomally normal embryos.
Oocyte-mediated somatic cell haploidization is a process in which a diploid cell halves its chromosomal content by segregating its homologue within the ooplasm. Replacing the donor oocyte nucleus with a patient's female diploid somatic nucleus can generate patient-genotyped oocytes. Insemination of these resulting constructs enables their activation and induces a reductive meiotic division, haploidizing the diploid female donor cell that can subsequently support syngamy with the male genome and create a zygote. So far, experimental data for this method have been limited and have not consistently proven the generation of chromosomally normal embryos. Overall, we achieved reconstruction of murine oocytes with a micromanipulation survival rate of 56.5%, and a correct haploidization and fertilization rate of 31.2%, resulting in a 12.7% blastocyst rate. Time-lapse analysis revealed that reconstructed embryos underwent a timely polar body extrusion and pronuclear appearance followed by a satisfactory embryonic cleavage, comparable with the control. Whole genome sequencing of the analyzed embryos indicated that 27.3% (6/22) were properly diploid. Our findings suggest that diploid cell haploidization may be a feasible technique for creating functional gametes in mammals.
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