Trypanosoma cruzi Chagas (ATCC® 30013)

Strain Designations: Culbertson  /  Depositor: E Tobie  /  Biosafety Level: 2

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Strain Designations Culbertson
Application
Vector borne research
Biosafety Level 2

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Isolation Human, Brazil, 1926
Product Format frozen
Storage Conditions Frozen Cultures:
-70°C for 1 week; liquid N2 vapor for long term storage

Freeze-dried Cultures:
2-8°C

Live Cultures:
See Protocols section for handling information
Type Strain no
Comments Guanine aminohydrolase
Cytidine aminohydrolase activity
Host specificity of ribosomal DNA variation
Axenic cultivation
Medium ATCC® Medium 1029: LIT medium
ATCC® Medium 1012: Diphasic blood agar medium
Growth Conditions
Temperature: 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°C to 25°C. Observe the culture daily and transfer when numerous trophozoites are observed.
Mycoplasma Unknown
Name of Depositor E Tobie
Special Collection NCRR Contract
Chain of Custody
ATCC <-- LS Diamond <-- E Tobie <-- JT Culbertson <-- London School Hyg. Trop. Med. <-- . . . <-- Instituto Oswaldo Cruz
References

J. Parasitol. 28: 155-158, 1942.

Diamond LS. Improved method for the monoxenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae with trypanosomatids. J. Parasitol. 54: 715-719, 1968. PubMed: 4319344

Nolan LL. Partial purification and characterization of guanine aminohydrolase from Trypanosoma cruzi. Curr. Microbiol. 11: 217-220, 1984.

Meyer H. The fine structure of the flagellum and kinetoplast-chondriome of Trypanosoma (Schizotrypanum) cruzi in tissue culture. J. Protozool. 15: 614-621, 1958.

Clark CG. Axenic Cultivation of Entamoeba dispar Brumpt 1925, Entamoeba insolita Geiman and Wichterman 1937 and Entamoeba ranarum Grassi 1879. J. Eukaryot. Microbiol. 42: 590-593, 1995. PubMed: 7581333

Iralu V. Trypanosoma cruzi cultivation in Trypticase Soy Broth with hemin and serum. J. Parasitol. 53: 653-654, 1967. PubMed: 6026862

Kidder GW. Characteristics of cytidine aminohydrolase activity in Trypanosoma cruzi and Crithidia fasciculata. J. Protozool. 31: 298-300, 1984. PubMed: 6381719

Clark CG, Pung OJ. Host specificity of ribosomal DNA variation in sylvatic Trypanosoma cruzi from North America. Mol. Biochem. Parasitol. 66: 175-179, 1994. PubMed: 7984184

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