title: Functional Human Oocytes Generated by Transfer of Polar Body Genomes.
abstract: 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.
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Ma
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA; Bioinformatics Program, University of California at San Diego, La Jolla, CA 92093, USA.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: O'Neil
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Nuria
authors.initials: N
authors.lastname: Marti Gutierrez
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors.forename: Manoj
authors.initials: M
authors.lastname: Hariharan
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors.forename: Zhuzhu Z
authors.initials: ZZ
authors.lastname: Zhang
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA; Bioinformatics Program, University of California at San Diego, La Jolla, CA 92093, USA.
authors.forename: Yupeng
authors.initials: Y
authors.lastname: He
authors.affiliation: IviGen Los Angeles, Torrance, CA 90501, USA.
authors.forename: Cengiz
authors.initials: C
authors.lastname: Cinnioglu
authors.affiliation: IviGen Los Angeles, Torrance, CA 90501, USA.
authors.forename: Refik
authors.initials: R
authors.lastname: Kayali
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Eunju
authors.initials: E
authors.lastname: Kang
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Yeonmi
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Tomonari
authors.initials: T
authors.lastname: Hayama
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Koski
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors.forename: Joseph
authors.initials: J
authors.lastname: Nery
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors.forename: Rosa
authors.initials: R
authors.lastname: Castanon
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Tippner-Hedges
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Riffat
authors.initials: R
authors.lastname: Ahmed
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: Van Dyken
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Molecular and Medical Genetics, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Olson
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: David
authors.initials: D
authors.lastname: Battaglia
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Lee
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Diana H
authors.initials: DH
authors.lastname: Wu
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Paula
authors.initials: P
authors.lastname: Amato
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Don P
authors.initials: DP
authors.lastname: Wolf
authors.affiliation: Genomic Analysis Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA; Howard Hughes Medical Institute, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA. Electronic address: ecker@salk.edu.
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Ecker
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, Portland, OR 97239, USA; Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR 97239, USA. Electronic address: mitalipo@ohsu.edu.
authors.forename: Shoukhrat
authors.initials: S
authors.lastname: Mitalipov
mesh_terms: D000328:Adult; D001755:Blastocyst / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics; D044127:Epigenesis, Genetic; D005260:Female; D005307:Fertilization in Vitro; D020869:Gene Expression Profiling; D015894:Genome, Human; D042822:Genomic Instability; D000066449:Human Embryonic Stem Cells / Q000378:metabolism; D006801:Humans; D008297:Male; D008677:Metaphase; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000378:metabolism*; D011003:Ploidies; D059705:Polar Bodies / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D013094:Spermatozoa / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism; D014158:Transcription, Genetic
keywords: haploid; infertility treatment; meiosis; mitochondrial replacement therapy; polar body nuclear transfer; reconstructed human oocytes