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作 者:燕海峰[1,2] 文平[2] 邓缘[2] 胡雄贵[2] P.Trefil 柳小春[1]
机构地区:[1]湖南农业大学动物科技学院,湖南长沙410128 [2]湖南省畜牧兽医研究所,湖南长沙410131 [3]捷克生物医药研究所
出 处:《激光生物学报》2008年第6期834-839,共6页Acta Laser Biology Sinica
基 金:畜禽种质资源标准化整理;整合及共享试点(2005DKA21101);中捷政府间科技合作项目(中方37-10捷方1P05ME722);湖南省科技攻关项目(2007WK3028)
摘 要:种蛋开窗对禽胚实施操作的技术在转基因禽类、利用多能细胞保存禽类种质资源以及禽胚实验模型等领域的研究具有重要价值。以家鸡新产蛋(含有禽胚胎干细胞)为材料,研究出了种蛋开窗切块自动复位技术,并在此基础上以接近禽胚实施操作为目的,进行了不同封口方法、不同开窗位置和添加液体排除因开窗形成的空气泡等对种蛋孵化率影响的实验。结果表明:种蛋赤道面开窗不同封口方式对孵化率影响的差异不显著,最高孵化率可达到65%;赤道面开窗,通过添加液体排除空气泡的影响不会提高孵化率,空气泡的大小与孵化率没有明显相关性;种蛋气室端开窗(针刺透内壳膜操作胚胎)方式,比赤道面开窗方式的孵化率明显提高,两种方法能见胚率均在90%以上;采用以上技术对种蛋开窗、找胚和封口,制作效率在每人每小时30个以上。本文系统地研究和报道了种蛋开窗操作禽胚简单而实用的技术,有助于解决目前种蛋开窗技术复杂且孵化率低的困境。Fertile egg windowing to access bird embryo is a useful technique in transgenic birds, conserving bird genetic resources with pluripotent stem cells as well as experimental embryo model fields. We invented a technique using newly laid fertile eggs, which the cover cut from the eggshell could recover automatically on the window. Based on this technique several factors affecting the hatchability of the windowed eggs ( HWE ) were investigated, such as different seal method, different windowing spot on the eggshell and adding liquid to drive air bubbles within the windowed eggs. It was indicated that the seal methods here did not remarkably affect the HWE and the highest results got up to 65 %. It seemed as if that adding liquid to move the air bubbles did not help to increase the HWE, and the volume of the bladder has also no visible effect on the HWE ; The HWE of the method, which windowed on the blunt end of the eggs ( access to the embryo by stinging through the inner membrane) was much higher than that of windowed on equatorial plane, and the rate of both methods to find the embryos was 90 % ; The efficiency was 30 eggs for one person one hour using the above methods to window, find embryo and recover the eggs. The technique reported here was very efficiency and practical, and it would do great helps to the research on transgenic poultry as well as relative fields.
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