The possibilities of optical laser technologies in cell engineering

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

doi:10.1134/s0012496609060295

Cited by 5 publications

(4 citation statements)

References 13 publications

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“…First results about functional enucleation of mammalian oocytes by laser irradiation of the metaphase plate were obtained in our experiments in mice [2] and repeated later on in pigs [3]. Furthermore, we showed that is possible to perform the entire mammalian cloning procedure at all stages of nuclear transplantation only with the use of laser [4].…”

Section: Introductionsupporting

confidence: 55%

“…Laser fusion of blastomeres at the two cell stage is a reliable technique for tetraploid embryo production and tetraploidy of all cells of laser created embryos was strongly proved by karyotyping as we reported [2]. Laser may be also used for the fusion donor karyoplasts with enucleated cytoplasts in somatic cell nuclear transfer (SCNT) as we demonstrated [4] and for increasing the oocyte volume after SCNT. Laser based methods may as be very efficient tools for novel developments in assisted reproductive technologies in humans e.g.…”

Section: Discussionmentioning

confidence: 82%

“…Later, the IR laser mediated fusion of large blastomeres of parthenogenetic two-cell porcine embryos was demonstrated [18]. We also reported, for the first time, successful laser fusion of mouse oocytes with somatic cumulus cells [4] where we used the laser with the same parameters as laid down by [2].…”

Section: Introductionmentioning

confidence: 89%

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Laser Fusion of Mouse Embryonic Cells and Intra-Embryonic Fusion of Blastomeres without Affecting the Embryo Integrity

Krivokharchenko

Karmenyan²,

Sarkisov

et al. 2012

PLoS ONE

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Manipulation with early mammalian embryos is the one of the most important approach to study preimplantation development. Artificial cell fusion is a research tool for various biotechnological experiments. However, the existing methods have various disadvantages, first of them impossibility to fuse selected cells within multicellular structures like mammalian preimplantation embryos. In our experiments we have successfully used high repetition rate picosecond near infrared laser beam for fusion of pairs of oocytes and oocytes with blastomeres. Fused cells looked morphologically normal and keep their ability for further divisions in vitro. We also fused two or three blastomeres inside four-cell mouse embryos. The presence of one, two or three nuclei in different blastomeres of the same early preimplantation mouse embryo was confirmed under UV-light after staining of DNA with the vital dye Hoechst-33342. The most of established embryos demonstrated high viability and developed in vitro to the blastocyst stage. We demonstrated for the first time the use of laser beam for the fusion of various embryonic cells of different size and of two or three blastomeres inside of four-cell mouse embryos without affecting the embryo’s integrity and viability. These embryos with blastomeres of various ploidy maybe unique model for numerous purposes. Thus, we propose laser optical manipulation as a new tool for investigation of fundamental mechanisms of mammalian development.

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Section: Introductionsupporting

confidence: 55%

Section: Discussionmentioning

confidence: 82%

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Laser Fusion of Mouse Embryonic Cells and Intra-Embryonic Fusion of Blastomeres without Affecting the Embryo Integrity

Krivokharchenko

Karmenyan²,

Sarkisov

et al. 2012

PLoS ONE

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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%

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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Femtosecond laser-induced blastomere fusion results in embryo tetraploidy by common metaphase plate formation

Osychenko

Zalessky

Astafiev

et al. 2020

Experimental Cell Research

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The possibilities of optical laser technologies in cell engineering

Cited by 5 publications

References 13 publications

Laser Fusion of Mouse Embryonic Cells and Intra-Embryonic Fusion of Blastomeres without Affecting the Embryo Integrity

Laser Fusion of Mouse Embryonic Cells and Intra-Embryonic Fusion of Blastomeres without Affecting the Embryo Integrity

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

Femtosecond laser-induced blastomere fusion results in embryo tetraploidy by common metaphase plate formation

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