Tissues from mouse, rat, and chick embryos have been cultured succesfully in three-dimensional collagen gels.
1. Dissect embryos from the decidua, or chick embryos from the eggshell into ice-cold L15-air (1). Dissect embryos away from membranes and adhering yolk, and assess developmental stage.
2. To isolate a specific portion of tissue, first dissect out that tissue surrounded by adjacent tissues, using either electrolytically sharpened tungsten needles or small Vannas scissors. Treatment with an enzyme such as Dispase, frequently is necessary to free the required explant from adjacent tissues. Place the tissue in a small volume (approx 1 mL) of Dispase at room temperature for 5-15 min. Once the reaction has occurred (4), remove tissues into cold L15-air (5) with a drop of serum included (6), and allow to rest on ice for approx 15-30 min before beginning to dissect out the appropriate portion of tissue using tungsten needles. Collect tissues into L15-air with a drop of serum, and keep on ice.
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