Developing chick embryos are a classical research tool in developmental biology. The whole embryo culture technique can be applied to various fields, such as embryo manipulation, toxicology, tumorigenesis, and basic research in regenerative medicine. When used for the generation of transgenic chickens, a high hatchability of genetically engineered embryos is essential to support normal embryonic development during culture. In this study, calcium carbonate, which is the main component of eggshells, was added as a calcium source in shell-less chick embryo cultures using a transparent plastic film as a culture vessel. In the absence of a calcium source in the shell-less culture system, embryogenesis ceased during culture, resulting in failed embryonic hatching. We found that the direct addition of calcium carbonate to the chorioallantoic membrane of the developing embryo was effective for the hatching of cultured chick embryos. The amount, timing, and location of calcium carbonate addition were investigated to maximize the hatchability of cultured embryos. Starting from the time of calcium carbonate supplementation, >40% hatchability was obtained with the optimal condition. This established method of shell-less chick embryo culture provides a useful tool in basic and applied fields of chick embryo manipulation.

发育中的雏鸡胚胎是发育生物学中的经典研究工具。整个胚胎培养技术可应用于各种领域，例如胚胎操作，毒理学，肿瘤发生和再生医学的基础研究。当用于产生转基因鸡时，转基因胚胎的高孵化率对于支持培养期间正常的胚胎发育至关重要。在这项研究中，使用透明塑料膜作为培养容器，在无壳鸡胚培养物中添加了作为蛋壳主要成分的碳酸钙作为钙源。在无壳培养系统中缺少钙源的情况下，在培养过程中胚胎发生停止，导致胚胎孵化失败。我们发现将碳酸钙直接添加到发育中的胚胎的绒膜尿囊膜对于孵化培养的雏鸡胚胎是有效的。研究了碳酸钙添加的量，时间和位置，以最大化培养的胚胎的孵化率。从添加碳酸钙开始，在最佳条件下可获得> 40％的孵化率。这种建立的无壳鸡胚培养方法为鸡胚处理的基础和应用领域提供了有用的工具。在最佳条件下可获得40％的孵化率。这种建立的无壳鸡胚培养方法为鸡胚处理的基础和应用领域提供了有用的工具。在最佳条件下可获得40％的孵化率。这种建立的无壳鸡胚培养方法为鸡胚处理的基础和应用领域提供了有用的工具。