Tetrahymena malaccensis Simon et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Handling of Test Tube CulturesThis strain is routinely shipped as a growing culture in a glass 16 x 125 mm screw-capped test tube. The volume of the cell suspension is approximately 5 ml. When the culture arrives remove it promptly from the shipping container. Do not store the culture at refrigeration temperatures before handling. To assure viability, immediately loosen the test tube cap and incubate upright at 25°C for at least one hour before observing the culture. There should be numerous active trophozoites in suspension. If the numbers are low the culture may have been exposed to temperature extremes in transit. Regardless of the state of the culture, aseptically transfer a 0.5 ml aliquot to a 16 x 125 mm screw-capped test tube containing 5 ml of sterilized ATCC medium 357. Incubate the parent and daughter cultures upright with the caps on loosely at 25°C
Routine Short-Term Maintenance of Culture:
Routine Long-Term Maintenance of Culture:
RM-9 Media for cryopreservation of Tetrahymena
Proteose Peptone (Difco 0120), 5.0 g
Tryptone, 5.0 g
K2HPO4, 0.2 g
Glucose, 1.0 g
Liver extract, 0.1 g
Glass distilled water, 1.0 L
Dissolve components in glass distilled H2O and autoclave.
Dryl's Salt Solution
0.1 M NaH2PO4 . 3H20, 10.0 ml
0.1 M Na2HPO4 . 7H20, 10.0 ml
0.1 M Sodium citrate . 2H20, 15.0 ml
0.1 M CaCl2 . 2H20, 15.0 ml
Distilled water, 950.0 ml
Add the first 3 components to the distilled H2O and mix thoroughly.
Add the CaC12 solution and mix thoroughly.
(Adding the solutions in the order indicated will avoid the precipitation of Ca salts.)
Alternative Thawing Procedure
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Simon EM, et al. New wild Tetrahymena from Southeast Asia, China, and North America, including T. malaccensis, T. asiatica, T. nanneyi, T. caudata, and T. silvana n. spp.. J. Protozool. 32: 183-189, 1985. PubMed: 3989748