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Attempt to produce nuclear transferred primordial germ cells using electrofusion in domestic chicken
Authors:Takeo MINEMATSU  Atsushi TAJIMA  Yukio KANAI
Institution:Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba-shi, Japan
Abstract:As a step to develop a somatic nuclear transfer technique for avian species, an attempt to produce somatic nuclear transferred primordial germ cells (PGC) in the domestic chicken was carried out. Primordial germ cells and embryonic blood cells (EBC) were collected from 2‐day‐old embryos and the nuclei were transferred from EBC into PGC by electrofusion. The most efficient pearl chain was developed when a 350‐V/cm AC field was applied for 60 s. Cell fusion between PGC and EBC was most effective when 4‐kV/cm DC pulses, 60 µs pulse width, were applied three times to a cell suspension dispersed in 0.2 or 0.25 mol/L saccharose solution. The present results provide basic information for the production of somatic cell nuclear transferred chickens using PGC as the nuclear recipient.
Keywords:chicken  electrofusion  nuclear transfer  primordial germ cells
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