The use of laser for obtaining recipient cytoplasts for mammalian nuclear transfer

Shakhbazyan, A. K.; Sviridova-Chailakhyan, T. A.; Karmenyan, A. K.; Krivokharchenko, Alexander; Chiou, Arthur; Chailakhyan, L. M.

doi:10.1134/s001249660905024x

Cited by 5 publications

(3 citation statements)

References 12 publications

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“…Inactivation of the oocyte metaphase plate using a near-infrared picosecond or femtosecond laser is an efficient and accurate method to prepare recipient cytoplasts. 1 – 3 A recipient cytoplast (an oocyte without chromosomes) is required for animal cloning and assisted reproductive technologies in humans, viz., for mitochondrial replacement therapy. 4 , 5 Removal of chromosomes lined up on the metaphase plate of the oocyte is commonly called enucleation.…”

Section: Introductionmentioning

confidence: 99%

Stain-free enucleation of mouse and human oocytes with a 1033 nm femtosecond laser

Osychenko,

Zalessky,

Bachurin

et al. 2024

J. Biomed. Opt.

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Results: We demonstrated a fundamental possibility of inactivating the metaphase plate in mouse and human oocytes by 1033 nm femtosecond laser radiation without applying a fluorescent dye. Irradiation of the spindle area, visualized by polarized light microscopy, resulted in partly or complete metaphase plate destruction but avoided the microtubules impairment. After the metaphase plate elimination, the spindle reorganized, however, it was not a complete depolymerization.Conclusions: This method of recipient cytoplast preparation is expected to be useful for animal cloning and assisted reproductive technologies.

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Section: Introductionmentioning

confidence: 99%

Stain-free enucleation of mouse and human oocytes with a 1033 nm femtosecond laser

Osychenko,

Zalessky,

Bachurin

et al. 2024

J. Biomed. Opt.

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“…The technology of cell fusion is used to obtain hybridomas 1 in terms of reprogramming and transforming the cell pathway, as well as for gene transfer and genetic therapy. 2 Moreover, laser nanosurgery approach can be successfully used to perform reproductive and therapeutic cloning, 3,4 including the tetraploid embryo application. 5 Femtosecond cell fusion is one of the innovative techniques, which can be applied to tetraploid embryo production by means of blastomere fusion.…”

mentioning

confidence: 99%

“…(2) how does the gas-vapor bubble affect the embryo development and how does the laser irradiation dose affect the probability of blastomere fusion and destruction, as well as embryo viability and quality? (3) what is the tetraploidization mechanism after the femtosecond-laser fusion? and (4) does the femtosecond-laser exposure affect the embryonic "clocks" in fusion, nonfusion, and half-destruction events?…”

mentioning

confidence: 99%

Femtosecond laser surgery of two-cell mouse embryos: effect on viability, development, and tetraploidization

et al. 2017

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The effect of the laser pulse energy and total expose of the energy incident on the embryo blastomere fusion probability was investigated. The probability of the four different events after laser pulse was determined: the fusion of two blastomeres with the following formation of tetraploid embryo, the destruction of the first blastomere occurs, the second blastomere conservation remains intact, the destruction and the death of both cells; two blastomeres were not fused, and no morphological changes occurred. We report on viability and quality of the embryo after laser surgery as a function of the laser energy incident. To characterize embryo quality, the probability of the blastocyst stage achievement was estimated and the blastocyst cells number was calculated. Blastocoel formation is the only event of morphogenesis in the preimplantation development of mammals, so we assumed it as an indicator of the time of embryonic "clocks" and observed it among fused and control embryos. The blastocoel formation time is the same for fused and control embryos. It indicates that embryo clocks were not affected due to blastomere fusion. Thus, the analysis of the fluorescence microscopic images of nuclei in the fused embryo revealed that nuclei fusion does not occur after blastomere fusion.

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Suppressing mosaicism by Au nanowire injector-driven direct delivery of plasmids into mouse embryos

et al. 2017

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The use of laser for obtaining recipient cytoplasts for mammalian nuclear transfer

Cited by 5 publications

References 12 publications

Stain-free enucleation of mouse and human oocytes with a 1033 nm femtosecond laser

Stain-free enucleation of mouse and human oocytes with a 1033 nm femtosecond laser

Femtosecond laser surgery of two-cell mouse embryos: effect on viability, development, and tetraploidization

Suppressing mosaicism by Au nanowire injector-driven direct delivery of plasmids into mouse embryos

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