A Simplified Tissue Culture Technic

Abstract

In a single chicken embryo, we find all that is required for preparing tissue cultures: the ingredients of the culture medium as well as the explant. Salt-glucose mixtures are replaced by amniotic fluid from the same embryo from which the tissues are obtained. An egg shell sector about 1 1/4 inches in diameter is cut out of the large egg pole on the 8-9th day of incubation, and the contents are poured cautiously into a Petri dish so as to avoid bursting the very delicate amniotic membrane. The membrane is then punched with a thin, sharp cannula and 2-3 cc of amniotic fluid are aspirated. The shell sector should not be larger than indicated, otherwise the amniotic membrane may be easily damaged. Only completely transparent fluid should be used; turbid fluids are mixtures of yolk and amniotic fluid and will inhibit growth. Tissue for explants is then cut out and the remaining embryonic tissue provides the embryonic juice. The culture medium consists of embryonic juice and amniotic fluid 1:1. The excess of the amniotic fluid may be used for all purposes where saline or other salt mixtures are usually used, e.g., washing of tissue, etc. The growth of cultures in this medium is not inferior to the growth of cultures in other fluid media. The cells are as normal in appearance as cells from a plasma culture (Fig. 1). Amniotic fluid not only replaces but also, in many regards, surpasses Tyrode's and other salt-glucose solutions in that it does not require sterilizing, filtering, or pH-adjusting. This liquid medium method may be recommended for cultures where the primary medium is to be removed for experimental purposes and replaced by other media.