Chapman S C, Collignon J, Schoenwolf G C, Lumsden A
MRC Centre for Developmental Neurobiology, King's College London, Guy's Hospital, London, United Kingdom.
Dev Dyn. 2001 Mar;220(3):284-9. doi: 10.1002/1097-0177(20010301)220:3<284::AID-DVDY1102>3.0.CO;2-5.
We describe a simple method of chick whole-embryo culture, which uses a filter paper carrier to hold the early blastoderm and vitelline membranes under tension while the embryo grows on a substratum of agar-albumen. This is a quick and efficient means of setting up cultures of chick embryos beginning at pre-primitive streak stages to stage 10 (stages X--XIV, Eyal-Giladi and Kochav [1976] Dev Biol 49:321-337; stages 1--10, Hamburger and Hamilton [1951] J Morphol 88:49--92). This is an improvement on the original method of New, which used a glass ring and watch glass (New [1955] Exp Morphol 3:320--331). Our modification of New's method, which we call EC (Early Chick, pronounced EASY) culture, facilitates several manipulations in early chick embryos, including microsurgery, grafting, bead implantation, microinjection, and electroporation. Using the EC method, embryos at stage 8 and older can be readily cultured either dorsal-side up (in contrast to New's method) or ventral-side up, as desired; embryos younger than stage 8 can be culture only ventral-side up (as with New's method). We also discuss some alternative methods for setting up these cultures.
我们描述了一种简单的鸡胚全胚胎培养方法，该方法使用滤纸载体在胚胎在琼脂 - 蛋白培养基上生长时，将早期胚盘和卵黄膜保持在张力状态。这是一种快速有效的方法，用于建立从原条前期到第10阶段（X - XIV阶段，Eyal - Giladi和Kochav [1976] 《发育生物学》49:321 - 337；第1 - 10阶段，Hamburger和Hamilton [1951] 《形态学杂志》88:49 - 92）的鸡胚培养。这是对New的原始方法的改进，后者使用玻璃环和表玻璃（New [1955] 《实验形态学》3:320 - 331）。我们对New方法的改进，我们称之为EC（早期鸡胚，发音为EASY）培养，便于对早期鸡胚进行多种操作，包括显微手术、移植、珠子植入、显微注射和电穿孔。使用EC方法，第8阶段及以上的胚胎可以根据需要很容易地以背侧向上（与New的方法相反）或腹侧向上的方式培养；小于第8阶段的胚胎只能以腹侧向上的方式培养（与New的方法一样）。我们还讨论了建立这些培养的一些替代方法。
