Ref ID : 3860
Kenneth W. Dobra, Eugene W. McArdle, and Charles F. Ehret; Growth Kinetics of Three Species of Tetrahymena on Solid Agar. J.Protozool. 27(2):226-230, 1980
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A nutrient-agar method without liquid overlay has been developed for cultivation of ciliates. Three species of Tetrahymena-T. pyriformis strain W, T. rostrata strain UNI, and T. vorax strain V2S, representing the 3 main groups of Tetrahymena species, were used; however the method should apply to other ciliates. Growth on the surface of the agar was facilitated by an optimal surface-to volume ratio yielding a high density of ciliates (5.8x10E5 cells/ml for T. pyriformis at 25 degrees C) and short generation times (3 hr for T. pyriformis at 30 degrees C). At the highest density achieved, the cells became irregularly hexagonal and formed a monolayer "tissue" on the agar. Ciliates grown on agar were like those in liquid culture, typical oral ciliature, food-vacuole formation, and typical cortical patterns being retained. Advantages of this method include high cell density, easy recovery, and optimal O2 supply. The organisms can also be cultivated on the surface of sterile cellulose-nitrate filters, facilitating in situ fixation and staining as well as transfer into different media by transfer of filters with cells, without prior centrifugation and resuspension.