doi: 10.15389/agrobiology.2013.2.46eng
UDC 636.2:576.315.086.862:591.39
OPTIMIZATION OF PARAMETERS OF THE OOCYTE ENUCLEATION AND FUSION WITH THE SOMATIC CELL DURING PRODUCTION OF MAMMALIAN CLONED EMBRYOS
G.N. Singina1, A.V. Lopukhov1, N.A. Zinovieva1, E.V. Shapkanova2, A.A. Puzik3
1All-Russian Research Institute of Animal Husbandry, Russian Academy of Agricultural Sciences,
pos. Dubrovitsy, Moscow Province, 142132 Russia,
e-mail: g_singina@mail.ru, vubi_myaso@mail.ru, n_zinovieva@mail.ru;
2Velikiye Luki State Agricultural Academy,
Velikiye Luki, Pskov Province, 182100 Russia,
e-mail: lenok150185@mail.ru;
3Smolensk Research Institute of Agriculture, Russian Academy of Agricultural Sciences,
21, ul. Nakhimova,Smolensk, 214025 Russia,
e-mail: smniish@yandex.ru
Received January 15, 2013
Parameters of the oocyte enucleation and fusion with somatic cell for producing of cattle cloned cetohybrids were studied. It was established that the electrofusion efficiency depends on strength, duration and set number of electrical pulse. The optimal parameters for Multiporator (Eppendorf, Germany) were determined: two consistent pulses DC electrical field each of 30 V field strength and 15 msec duration followed by the pulse of AC electrical field of 5 V and 10 sec duration. It was found that the first polar body serving as a marker for the chromosome position at «blind» enucleation must not deviate from the chromosome location by more than 10 degrees.
Keywords: in vitro culture, oocyte, cattle nuclear transfer, electrofusion, enucleation.
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