Functional Human Oocytes Generated by Transfer of Polar Body Genomes

• Published in: Cell stem cell 2017-01, Vol.20 (1), p.112-119
• Main Authors: Ma, Hong, O’Neil, Ryan C., Marti Gutierrez, Nuria, Hariharan, Manoj, Zhang, Zhuzhu Z., He, Yupeng, Cinnioglu, Cengiz, Kayali, Refik, Kang, Eunju, Lee, Yeonmi, Hayama, Tomonari, Koski, Amy, Nery, Joseph, Castanon, Rosa, Tippner-Hedges, Rebecca, Ahmed, Riffat, Van Dyken, Crystal, Li, Ying, Olson, Susan, Battaglia, David, Lee, David M., Wu, Diana H., Amato, Paula, Wolf, Don P., Ecker, Joseph R., Mitalipov, Shoukhrat
• Format: Article
• Language: English
• Subjects: Adult; Blastocyst - metabolism; DNA Methylation - genetics; Embryonic Development - genetics; Epigenesis, Genetic; Female; Fertilization in Vitro; Gene Expression Profiling; Genome, Human; Genomic Instability; haploid; Human Embryonic Stem Cells - metabolism; Humans; infertility treatment; Male; meiosis; Metaphase; mitochondrial replacement therapy; Nuclear Transfer Techniques; Oocytes - metabolism; Ploidies; Polar Bodies - metabolism; polar body nuclear transfer; reconstructed human oocytes; Sequence Analysis, RNA; Spermatozoa - metabolism; Spindle Apparatus - metabolism; Transcription, Genetic
• ISSN: 1934-5909; 1875-9777
• DOI: 10.1016/j.stem.2016.10.001

Oocyte defects lie at the heart of some forms of infertility and could potentially be addressed therapeutically by alternative routes for oocyte formation. Here, we describe the generation of functional human oocytes following nuclear transfer of first polar body (PB1) genomes from metaphase II (MII) oocytes into enucleated donor MII cytoplasm (PBNT). The reconstructed oocytes supported the formation of de novo meiotic spindles and, after fertilization with sperm, meiosis completion and formation of normal diploid zygotes. While PBNT zygotes developed to blastocysts less frequently (42%) than controls (75%), genome-wide genetic, epigenetic, and transcriptional analyses of PBNT and control ESCs indicated comparable numbers of structural variations and markedly similar DNA methylation and transcriptome profiles. We conclude that rescue of PB1 genetic material via introduction into donor cytoplasm may offer a source of oocytes for infertility treatment or mitochondrial replacement therapy for mtDNA disease. [Display omitted] •Metaphase II oocytes can be reconstructed by polar body nuclear transfer (PBNT)•Reconstructed PBNT oocytes complete meiosis after fertilization with sperm•PBNT-derived blastocysts can give rise to phenotypically normal hESC lines Ma et al. show regeneration of functional human oocytes through polar body transfer into enucleated oocyte cytoplasts. In addition to providing proof of principle for this process, the approach could be helpful clinically for some forms of infertility and genetic disease.