Trypanosoma cruzi Chagas (ATCC® 50824)

Organism: Trypanosoma cruzi Chagas  /  Depositor: JA Dvorak

Strain Designations SYLVIO-X10 CL7
Application
Vector borne research
Biosafety Level 2
Isolation Clone 7 derived from strain SYLVIO X10 (ATCC 50823)
Product Format frozen
Storage Conditions Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Medium Medium 1029: LIT medium
Growth Conditions
Temperature: 20°C to 25°C
Culture System: Axenic
Cryopreservation Harvest and Preservation
  1. Harvest cells from several cultures in very late logarithmic to early stationary phase of growth.  Vigorously agitate to suspend the cells.
  2. Aseptically transfer the cell suspension to 15 mL plastic centrifuge tubes.
  3. Centrifuge at ~800 x g for 5 min.
  4. While cells are centrifuging, prepare a 10% solution of DMSO in complete ATCC Medium 1029.  Cool on ice.
  5. Remove the supernatant and pool the cell pellets to the final volume desired with fresh growth medium.
  6. Combine the cell suspension with an equal volume of 10% DMSO cryoprotectant solution (prepared in step 4) to yield a final concentration of 5% DMSO.
  7. Dispense in 0.5 mL aliquots to 1.0-2.0 mL Nunc vials (special plastic vials for cryopreservation).
  8. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  At -40°C, plunge ampules into liquid nitrogen.  Alternatively, place the vials 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.).  
  9. Store ampules in a liquid nitrogen refrigerator until needed.
  10. To establish a culture from the frozen state, place a frozen ampule in a 35°C water bath just enough to cover the frozen material.  Allow the ampule to thaw completely (2-3 min).
  11. Immediately after thawing, aseptically remove the contents and transfer to a T-25 tissue culture flask containing 10 mL of fresh complete ATCC medium 1029.
  12. Screw the cap on tightly and incubate at 20-25°C.    Observe the culture daily and transfer when numerous trophozoites are observed.           
Name of Depositor JA Dvorak
Special Collection NCRR Contract
References

Dvorak JA, et al. Trypanosoma cruzi: flow cytometric analysis. I. Analysis of total DNA/organism by means of mithramycin-induced fluorescence. J. Protozool. 29: 430-437, 1982. PubMed: 6182288

Miyahira Y, Dvorak JA. Kinetoplastidae display naturally occurring ancillary DNA-containing structures. Mol. Biochem. Parasitol. 65: 339-349, 1994. PubMed: 7969274

Nozaki T, et al. Cellular and molecular biological analyses of nifurtimox resistance in Trypanosoma cruzi. Am. J. Trop. Med. Hyg. 55: 111-117, 1996. PubMed: 8702014

McDaniel JP, Dvorak JA. Identification, isolation, and characterization of naturally-occurring Trypanosoma cruzi variants. Mol. Biochem. Parasitol. 57: 213-222, 1993. PubMed: 8433713

Finley RW, Dvorak JA. Trypanosoma cruzi: analysis of the population dynamics of heterogeneous mixtures. J. Protozool. 34: 409-415, 1987. PubMed: 3323478

Nozaki T, Dvorak JA. Intraspecific diversity in the response of Trypanosoma cruzi to environmental stress. J. Parasitol. 79: 451-454, 1993. PubMed: 8501607

Postan M, et al. Studies of Trypanosoma cruzi clones in inbred mice. I. A comparison of the course of infection of C3H/HEN- mice with two clones isolated from a common source. Am. J. Trop. Med. Hyg. 32: 497-506, 1983. PubMed: 6407346