Trypanosoma theileri Laveran (ATCC® 30017)

Organism: Trypanosoma theileri Laveran  /  Depositor: EJ Soulsby

Permits and Restrictions

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Application
Vector borne research
Biosafety Level 1
Isolation
blood - animal
Product Format frozen
Type Strain no
Comments
This strain is useful for bovine trypanosomiasis and lymphocytosis parallel studies.
Medium ATCC® Medium 482: MPHM medium
Growth Conditions
Temperature: 35.0°C
Duration: axenic
Protocol: See general instructions for thawing and storage of frozen material before proceeding. Add thawed contents to a single 16 x 125 mm glass screw-capped test tube of the appropriate medium. Incubate the culture vertically with the cap screwed on tightly. It is essential to establish cultures initially in small volumes. Once established, the culture can be scaled up to larger volumes. Vigorously agitate the culture and aseptically transfer 0.1 ml of culture to a fresh tube of medium weekly.
Subcultivation
Protocol: See general instructions for thawing and storage of frozen material before proceeding. Add thawed contents to a single 16 x 125 mm glass screw-capped test tube of the appropriate medium. Incubate the culture vertically with the cap screwed on tightly. It is essential to establish cultures initially in small volumes. Once established, the culture can be scaled up to larger volumes. Vigorously agitate the culture and aseptically transfer 0.1 ml of culture to a fresh tube of medium weekly.
Cryopreservation

1.   Harvest cells from a culture which is at or near peak density by centrifugation at 1,300 g for 5 min.

2.   Adjust concentration of cells to 2 x 107/ml in fresh medium.

3.   While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium (broth).  The DMSO solution when first prepared will warm up due to chemical heat. The solution should be allowed to return to room temperature prior to use.

4.   Mix the cell preparation and the DMSO solution in equal portions. The final concentration will be 107 cells/ml and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no more than 15 min.

5.   Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

6.   Place the ampules in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  

7.   Store in either the vapor or liquid phase of a nitrogen refrigerator.

8.   To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after thawed.

9.   Immediately after thawing, do not leave in the water bath, aseptically transfer the contents of the ampule into a fresh tube of ATCC medium 807. 

10.          Incubate vertically at 35°C with the cap screwed on tightly.

11.          Maintain as described above. 

Name of Depositor EJ Soulsby
Special Collection ATCC
Chain of Custody
ATCC <<--EJ Soulsby<<--A. Sollod
References

Sollod AE, Soulsby EJ. Cultivation of Trypanosoma theileri at 37 C in partially defined media. J. Protozool. 15: 463-466, 1968. PubMed: 5703078

Hare WC, et alBovine trypanosomiasis and lymphocytosis parallel studiesIn: Hare WC, et alComparative leukemia research 1969No. 36BaselKargerpp. 504-507, 1970

Hare WC, Soulsby EJ. Stimulation of DNA synthesis in bovine leukocyte cultures by Trypanosoma theileri antigen. J. Parasitol. 55: 973-976, 1969. PubMed: 5391314

Splitter EJ, et al. Gel-diffusion and hemagglutination studies with Trypanosoma theileri. Exp. Parasitol. 20: 160-166, 1967. PubMed: 4962447

Splitter EJ, Soulsby EJ. Isolation and continuous cultivation of Trypanosoma theileri in media containing tissue culture fluids. Exp. Parasitol. 21: 137-148, 1967. PubMed: 4169719

dairy cow

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation