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New defined and semi-defined media for cultivation of the flagellate Trichomonas vaginalis.

Studies on the nutrition and physiology of Trichomonas vaginalis have been hampered by the lack of a defined synthetic medium in which to grow the organism. New synthetic media free of ill-defined supplements and serum which support good growth of T. vaginalis have been developed in this study. The first successful approach to a defined medium resulted from the modification of the tissue-culture medium CMRL 1066. Essential modifications included (1) lowering the redox potential of the medium by adding high levels of ascorbate and (2) modifying the buffer system. Subsequently, CMRL 1066 was replaced by two new media DL7 and DL8 which have greater flexibility in their formulation and permit improved serum-free growth. Serum was replaced by bovine serum albumin and cholesterol, together with either a glyceryl fatty acid ester or a defined mixture of fatty acids. Preliminary nutritional studies using the new media have indicated a requirement for saturated and unsaturated fatty acids and a sterol.

Animals↗

Primordial germ cells and oocytes of Branchiostoma virginiae (Cephalochordata, Acrania) are flagellated epithelial cells: relationship between epithelial and primary egg polarity.

Primordial germ cells (PGCs) are described from the gonad of c. 2 cm juvenile Branchiostoma virginiae; early oocytes (c. 10 microm) and enlarging, previtellogenic oocytes (c. 35 microm) are described from the ovary of c. 5 cm adults. The germinal epithelium of the juvenile gonad and adult ovary is composed of both germinal and somatic cells. In the juvenile, somatic cells retain contact with the basal lamina of the germinal epithelium though their perikarya may be displaced towards the lumen; the germinal epithelium is, therefore, a simple but pseudostratified epithelium. In the adult ovary, somatic cells may lose contact with the basal lamina and the epithelium appears to become stratified. PGCs and oocytes are identified as germ cells by the presence of nuage. PGCs and oocytes are polarised epithelial cells. They rest on a basal lamina, extend apically towards a lumen, form adhering junctions with neighbouring cells, and exhibit apical-basal polarity. PGCs and early oocytes have an apical flagellum with an associated basal body, accessory centriole, and one or more striated rootlet fibres. The flagellum is surrounded by a collar of microvilli. Once oocytes begin to enlarge and bulge basally into the connective tissue layer, the flagellum is lost, but the basal bodies and ciliary rootlets are present at the apex of 35 microm oocytes. Similarities of the oogenic pattern in cephalochordates and echinoderms indicate that the establishment of egg polarity in deuterostomes is influenced by the polarity of the germinal epithelium.

Animals↗