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Effect of Laser Optoperforation of the Zona Pellucida on Mouse Embryo Development in vitro

E. O. Zakharchenko1, A. D. Zalessky1,2, A. A. Osychenko1, A. S. Krivokharchenko1, A. K. Shakhbazyan1, A. V. Ryabova3, and V. A. Nadtochenko1,4,5*

1Semenov Institute of Chemical Physics, Russian Academy of Sciences, 119991 Moscow, Russia; E-mail: nadtochenko@gmail.com

2Moscow Institute of Physics and Technology, State University, 141700 Dolgoprudny, Moscow Region, Russia

3Prokhorov General Physics Institute, Russian Academy of Sciences, 119991 Moscow, Russia

4Institute of Problems of Chemical Physics, Russian Academy of Sciences, 142432 Chernogolovka, Moscow Region, Russia

5Lomonosov Moscow State University, Faculty of Chemistry, 119991 Moscow, Russia

* To whom correspondence should be addressed.

Received January 13, 2015; Revision received February 23, 2015
The effect of laser optical perforation of the zona pellucida on the viability and development of mouse embryos has been studied. Operations of zona pellucida thinning and single or double perforation were carried out on 2-cell embryo, morula, and blastocyst stages with a laser pulse (wavelength 1.48 µm, pulse duration 2 ms). Embryo development up to the blastocyst stage and hatching efficiency were statistically analyzed. It was found that 2-cell or morula stage embryo zona pellucida thinning or single perforation did not affect development to the blastocyst stage and number of hatched embryos, but it accelerated embryo hatching compared to control groups one day earlier in vitro. Double optoperforation on 2-cell embryo or morula stage did not significantly affect development to the blastocyst stage, but it strongly decreased the number of hatched embryos. Also, zona pellucida perforation at the blastocyst stage had a negative effect: hatching did not occur after this manipulation. Blastocyst cell number calculation after single zona pellucida perforation at 2-cell and morula stages showed that cell number of hatching or hatched blastocysts did not differ from the same control groups. This fact points out that the laser single optoperforation method is a useful and safe experimental tool that allows further manipulations within the zona pellucida.
KEY WORDS: biophotonics, photobiology, laser optoperforation, embryo, laser hatching

DOI: 10.1134/S0006297915060127