A Novel Shell-less Culture System for Chick Embryos Using a Plastic Film as Culture Vessels
A Novel Shell-less Culture System for Chick Embryos Using a Plastic Film as Culture Vessels
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DOI：
10.2141/jpsa.0130043
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发表时间：
2014-07-01
影响因子：
1.5
通讯作者：
Obara, Katsuya
中科院分区：
文献类型：
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作者：
Tahara, Yutaka;Obara, Katsuya
高孵化率鸡胚无壳培养技术的发展将有助于转基因鸡的高效生产、胚胎操作、组织工程和再生医学的基础研究。迄今为止，鸟类胚胎培养方法的研究包括使用窄窗蛋壳、代用蛋壳和使用透气膜的人工容器的全胚胎培养。然而，没有报道使用完全人工血管实现>50%的高孵化率。为了建立一种简便、高孵化率的鸡胚培养方法，我们对各种培养条件进行了研究，包括补钙和充氧的方法。在胚胎培养物中，胚胎在55-56小时孵育后转移到培养容器中，超过90%的胚胎存活到第17天，此时使用聚甲基戊烯膜作为具有乳酸钙和蒸馏水的培养容器。从第17天开始对存活的胚胎通纯氧，孵化率为57.1%（8/14）。因此，我们成功地实现了高孵化率与此方法在鸡胚培养使用人工血管。
The development of shell-less culture methods for bird embryos with high hatchability would be useful for the efficient generation of transgenic chickens, embryo manipulations, tissue engineering, and basic studies in regenerative medicine. To date, studies of culture methods for bird embryos include the whole embryo culture using narrow windowed eggshells, surrogate eggshells, and an artificial vessel using a gas-permeable membrane. However, there are no reports achieving high hatchability of >50% using completely artificial vessels. To establish a simple method for culturing chick embryos with high hatchability, we examined various culture conditions, including methods for calcium supplementation and oxygen aeration. In the embryo cultures where the embryos were transferred to the culture vessel after 55-56 h incubation, more than 90% of embryos survived until day 17 when a polymethylpentene film was used as a culture vessel with calcium lactate and distilled water supplementations. The aeration of pure oxygen to the surviving embryos from day 17 yielded a hatchability of 57.1% (8 out of 14). Thus, we successfully achieved a high hatchability with this method in chicken embryo culture using an artificial vessel.