| Popis: |
Publisher Summary This chapter discusses the culturing of rat pituitary explants. The aseptic removal of the rat or mouse pituitary is achieved by a series of successive steps, each requiring flaming of instruments or a new set of sterile instruments. The animal is killed by cervical dislocation or decapitation and is thoroughly wetted down with 70% alcohol to prevent hair and bacterial contamination of the area to be exposed. The skin over the skull is removed by making a small incision at the base of the neck just behind the occipital protuberance. The most convenient vessel for maintaining explants of the hypophyseal pars distalis is the presterilized plastic organ culture dish, which is used in conjunction with a triangular stainless steel grid. These plastic dishes are provided with an absorbent disk which should be aseptically moistened with 5 ml of sterile saline. Explants are prepared by cutting the pars distalis into small pieces with a pair of sterile scalpels, using opposed scalpel blades in a scissorlike fashion. It is important to size the explants fairly accurately because too large of an explant will develop central necrosis while too small of an explant will lose its histological identity. |
