Attempt to produce nuclear transferred primordial germ cells using electrofusion in domestic chicken
Attempt to produce nuclear transferred primordial germ cells using electrofusion in domestic chicken
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DOI：
10.1111/j.1740-0929.2004.00186.x
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发表时间：
2004-06
影响因子：
2
通讯作者：
T. Minematsu;A. Tajima;Y. Kanai
中科院分区：
文献类型：
--
作者：
T. Minematsu;A. Tajima;Y. Kanai
作为发展禽类体细胞核移植技术的一步，尝试在家鸡体细胞核移植原始生殖细胞(PGC)。从2日龄胚胎中采集原始生殖细胞和胚胎血细胞(EBC)，通过电融合的方法将EBC的细胞核移植到PGC中。当施加350V/cm的交流电场时，60 S的珠链最有效。当4kV/cm的直流脉冲、60µm的S脉冲作用于分散在0.2mol/L蔗糖溶液中的细胞悬液3次时，PGC与EBC的细胞融合效果最好。本研究结果为以PGC为核受体的体细胞核移植鸡的生产提供了基础资料。
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.