TY - JOUR
T1 - Towards germline gene therapy of inherited mitochondrial diseases
AU - Tachibana, Masahito
AU - Amato, Paula
AU - Sparman, Michelle
AU - Woodward, Joy
AU - Sanchis, Dario Melguizo
AU - Ma, Hong
AU - Gutierrez, Nuria Marti
AU - Tippner-Hedges, Rebecca
AU - Kang, Eunju
AU - Lee, Hyo Sang
AU - Ramsey, Cathy
AU - Masterson, Keith
AU - Battaglia, David
AU - Lee, David
AU - Wu, Diana
AU - Jensen, Jeffrey
AU - Patton, Phillip
AU - Gokhale, Sumita
AU - Stouffer, Richard
AU - Mitalipov, Shoukhrat
N1 - Funding Information:
Acknowledgements The authors would like to acknowledge the Oregon Health & Science University (OHSU) Embryonic Stem Cell Research Oversight Committee and the Institutional Review Board for providing oversight and guidance. We thank oocyte and sperm donors and staff at the Women’s Health Research Unit at the Center for Women’s Health, University Fertility Consultants and Reproductive Endocrinology & Infertility Division at the Department of Obstetrics & Gynecology of Oregon Health & Science University for their support and procurement of human gametes. The Division of Animal Resources, Surgery Team, Assisted Reproductive Technology & Embryonic Stem Cell Core, Endocrine Technology Core, Imaging & Morphology Core, Flow Cytometry Core and Molecular & Cellular Biology Core at the Oregon National Primate Research Center provided expertise and services for the nonhuman primate research. Hamilton ThorneInc.,donatedanXYClone laser systemfor thisstudy. Weare gratefulto W. Sanger and D. Zaleski for karyotyping services, C. Penedo for microsatellite analysis and J. Hennebold for consulting on metabolic assays. We are also indebted to A. Steele, R. Cervera Juanes and E. Wolff for their technical support. The human oocyte/embryo research was supported by grants from the OHSU Center for Women’s Health Circle of Giving and other OHSU institutional funds, as well as the Leducq Foundation. The nonhuman primate study was supported by grants from the National Institutes of Health HD063276, HD057121, HD059946, EY021214 and 8P51OD011092.
PY - 2013/1/31
Y1 - 2013/1/31
N2 - 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.
AB - 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.
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U2 - 10.1038/nature11647
DO - 10.1038/nature11647
M3 - Article
C2 - 23103867
AN - SCOPUS:84873088209
SN - 0028-0836
VL - 493
SP - 627
EP - 631
JO - Nature
JF - Nature
IS - 7434
ER -