A reliable technique of nuclear transplantation for immature mammalian oocytes
Autor: | Gianpiero D. Palermo, Zev Rosenwaks, Berrin Ergün, Takumi Takeuchi, Tian Hua Huang |
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Rok vydání: | 1999 |
Předmět: |
Nuclear Transfer Techniques
Enucleation Aneuploidy Biology Cell Fusion Andrology Mice Meiosis medicine Animals Metaphase Cellular Senescence Germinal vesicle Cell Cycle Rehabilitation Reproducibility of Results Obstetrics and Gynecology Anatomy medicine.disease Oocyte Transplantation medicine.anatomical_structure Reproductive Medicine Karyotyping Oocytes Female Cell aging |
Zdroj: | Human Reproduction. 14:1312-1317 |
ISSN: | 1460-2350 |
Popis: | Transplanting a germinal vesicle (GV) to another enucleated oocyte provides a possible way to avoid age-related aneuploidy in metaphase II (MII) oocytes from older women. This study was conducted to examine the efficiency of each step of nuclear transplantation as reflected in the survival and maturation capacity of immature mouse oocytes subjected to this procedure. GV stage oocytes were retrieved from unstimulated ovaries. A GV removed with a small amount of cytoplasm (karyoplast) was transferred subzonally into a previously enucleated oocyte, which was then exposed to direct current to promote fusion. Such reconstituted oocytes were placed in culture to allow maturation, and some that had extruded a first polar body were fixed and processed for chromosome analysis. Each step of nuclear transplantation - survival, enucleation, grafting, and reconstitution - was successful in >90%, with the overall efficiency of reconstitution being 80%. The observation of normal karyotypes confirmed that the procedure did not increase chromosomal aneuploidy. An electrolytic medium, revealed to be superior for the reconstitution procedure, also allowed haploidization of the transplanted nucleus. These findings suggest that this technique can be applied to study the effects of a 'younger' woman's ooplasm on the disjunction of an 'older' woman's chromosomes during meiosis I. |
Databáze: | OpenAIRE |
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