title: Towards germline gene therapy of inherited mitochondrial diseases.
abstract: Mutations in mitochondrial DNA (mtDNA) are associated with severe human diseases and are maternally inherited through the egg's cytoplasm. Here we investigated the feasibility of mtDNA replacement in human oocytes by spindle transfer (ST; also called spindle-chromosomal complex transfer). Of 106 human oocytes donated for research, 65 were subjected to reciprocal ST and 33 served as controls. Fertilization rate in ST oocytes (73%) was similar to controls (75%); however, a significant portion of ST zygotes (52%) showed abnormal fertilization as determined by an irregular number of pronuclei. Among normally fertilized ST zygotes, blastocyst development (62%) and embryonic stem cell isolation (38%) rates were comparable to controls. All embryonic stem cell lines derived from ST zygotes had normal euploid karyotypes and contained exclusively donor mtDNA. The mtDNA can be efficiently replaced in human oocytes. Although some ST oocytes displayed abnormal fertilization, remaining embryos were capable of developing to blastocysts and producing embryonic stem cells similar to controls.
authors.affiliation: Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, Oregon Health & Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Masahito
authors.initials: M
authors.lastname: Tachibana
authors.forename: Paula
authors.initials: P
authors.lastname: Amato
authors.forename: Michelle
authors.initials: M
authors.lastname: Sparman
authors.forename: Joy
authors.initials: J
authors.lastname: Woodward
authors.forename: Dario Melguizo
authors.initials: DM
authors.lastname: Sanchis
authors.forename: Hong
authors.initials: H
authors.lastname: Ma
authors.forename: Nuria Marti
authors.initials: NM
authors.lastname: Gutierrez
authors.forename: Rebecca
authors.initials: R
authors.lastname: Tippner-Hedges
authors.forename: Eunju
authors.initials: E
authors.lastname: Kang
authors.forename: Hyo-Sang
authors.initials: HS
authors.lastname: Lee
authors.forename: Cathy
authors.initials: C
authors.lastname: Ramsey
authors.forename: Keith
authors.initials: K
authors.lastname: Masterson
authors.forename: David
authors.initials: D
authors.lastname: Battaglia
authors.forename: David
authors.initials: D
authors.lastname: Lee
authors.forename: Diana
authors.initials: D
authors.lastname: Wu
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Jensen
authors.forename: Phillip
authors.initials: P
authors.lastname: Patton
authors.forename: Sumita
authors.initials: S
authors.lastname: Gokhale
authors.forename: Richard
authors.initials: R
authors.lastname: Stouffer
authors.forename: Shoukhrat
authors.initials: S
authors.lastname: Mitalipov
mesh_terms: D000328:Adult; D000818:Animals; D002467:Cell Nucleus / Q000235:genetics; D015925:Cryopreservation; D003593:Cytoplasm / Q000235:genetics; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics; D004622:Embryo, Mammalian / Q000196:embryology; D053595:Embryonic Stem Cells / Q000166:cytology; D005260:Female; D005306:Fertilization; D015316:Genetic Therapy; D006801:Humans; D008253:Macaca mulatta / Q000235:genetics / Q000254:growth & development; D018895:Microsatellite Repeats / Q000235:genetics; D028361:Mitochondrial Diseases / Q000235:genetics* / Q000628:therapy*; D053652:Nuclear Transfer Techniques / Q000592:standards*; D009865:Oocytes / Q000166:cytology; D011247:Pregnancy; D055815:Young Adult; D015053:Zygote / Q000166:cytology / Q000473:pathology
chemical_list: D004272:DNA, Mitochondrial