The development of shellless 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. Studies on culture methods for avian embryos include whole embryo culture using eggshells with narrow windows, surrogate eggshells, and an artificial vessel using a gas-permeable membrane to achieve high hatchability above 50% using completely artificial vases. Thus, our group has established a simple method to culture chick embryos with high hatchability, we examined various culture conditions, including methods for calcium supplementation and oxygen aeration. In embryo cultures where the embryos were transferred to the culture vessel after 8 days of incubation, more than 90% of the embryos survived until day 12, when a polymethylpentene film was used as a culture vessel with sodium lactate supplements. calcium and distilled water. Pure oxygen aeration on surviving embryos from day 8 resulted in no hatchability until day 14. Only one egg survives for 14 days. Mostly, you die on day 10. They only survive 2 days after the first experiment where the egg was on the 8th day of development. Therefore, our laboratory has not successfully achieved high hatchability with this method in culturing chicken embryos using an artificial vessel. Say no to plagiarism. Get a tailor-made essay on "Why Violent Video Games Shouldn't Be Banned"? Get an original essay. Fertilized chicken eggs incubated in ambient air were examined at 8, 9, 10, 11, 12, 13, and 14 days of incubation and compared with eggs incubated in 60% oxygen. Eggs incubated in high-oxygen environments had lower diffusion capacities for carbon monoxide, lower rates of oxygen consumption per gram of embryo, heavier embryos, and more advanced embryonic morphological development than eggs incubated for the same time in ambient air. The lower rates of oxygen consumption per gram of embryo are consistent with an increased growth rate, as oxygen consumption (per gram of embryo) decreases with increasing weight; the lower diffusion capacities of eggs incubated in 60% oxygen are interpreted to be due to a reduced surface area of ​​the chorioallantoic capillaries, or an increased thickness of the diffusion barrier between the chick's blood and air, or both. eggshell on the process of shell calcium mobilization by the chorioallantoic membrane (CAM), chick embryos were maintained in long-term in vitro cultures without shells. The shellless embryos were severely calcium deficient and showed signs of delayed development and abnormal skeletal calcification. During development, calcium transport and calcium binding protein (CaBP) activities were decreased in the CAM of shellless embryos compared to those of control embryos developing in ovo. The levels of carbonic anhydrase activity expressed during development remained, however, similar. By a single radial CaBP immunodiffusion assay using a specific anti-CaBP antiserum, the level of immunoreactive CaBP was significantly increased in the CAM of shellless embryos. These studies indicate that the CAM of chick embryos cultured in shellless conditions is defective in calcium transport, possibly due to the expression of an inactive form of CaBP.