Supplemental calcium increases hatch rate but not hatchling mass of chick embryos in shell-less culture

A method is described for culturing 64-70 h-old chicken embryos and egg contents outside of the eggshell through to hatching. Cultured egg contents were suspended in polymethylpentene kitchen wrap (F.O.R. wrap; Riken Fabro) supported in polyvinyl chloride tripods. Tripods were incubated in Plexiglas environmental chambers which were rocked automatically through an angle of ±20°. The concentration of CO₂ was maintained at 2% throughout incubation, while that of O₂ was increased from ambient to 50%, and relative humidity was decreased from 90%-92% to 83%-84% at incubation Day 9. Cultured embryos not supplemented with calcium did not hatch. The Hatch rate increased when supplemental calcium L-lactate hydrate was increased between 250 and 350 mg. A maximal hatch rate of 54.8% was achieved when cultures were supplemented with 350 mg of calcium L-lactate hydrate and 3.5 ml of sterile water. Adding 400 or 450 mg of calcium L-lactate hydrate did not increase the hatch rate further. The mass of cultured hatchlings (including the retracted yolk) and yolk-free carcass wet and dry mass and length of the right third toe were significantly less than the corresponding parameters observed in hatchlings in ovo. No statistically significant differences in hatchling mass, yolk-free carcass wet or dry mass, or length of the right third toe were noted among cultured hatchlings supplemented with 250-450 mg of calcium L-lactate hydrate. Failure to completely absorb albumen was the most common abnormality observed in cultures which failed to hatch. The present technique allows a unique approach to study the physiology of the developing chicken embryo.