Trypanosoma cruzi Chagas (ATCC® 50791)

Strain Designations: M/HOM/AR/74/CA-I CL72  /  Depositor: JA Dvorak  /  Biosafety Level: 2

Strain Designations M/HOM/AR/74/CA-I CL72
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.

clone 72 derived from strain CA-I, originally isolated from a human male with chronic myocarditis, San Luis Province, Argentina, 1974, cloned by J. Dvorak, 1980
Product Format frozen
Type Strain no
flow cytometric analysis
growth kinetics of clones in liquid medium
Medium ATCC® Medium 1029: LIT medium
Growth Conditions
Temperature: 25.0°C
Duration: axenic

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 flask containing 10 ml ATCC medium 1029. 

10.          Incubate at 25°C with the cap screwed on tightly.

11.          Maintain as described above. 

Name of Depositor JA Dvorak
Special Collection NCRR Contract
Year of Origin 1974

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

Engel JC, et al. Trypanosoma cruzi: biological characterization of 19 clones derived from two chronic chagasic patients. I. Growth kinetics in liquid medium. J. Protozool. 29: 555-560, 1982. PubMed: 6816924

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

Engel JC, et al. Trypanosoma cruzi: biological characterization of clones derived from chronic chagasic patients. II. Quantitative analysis of the intracellular cycle. J. Protozool. 32: 80-83, 1985. PubMed: 3921699

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

Doyle PS, et al. Trypanosoma cruzi: quantification and analysis of the infectivity of cloned stocks. J. Protozool. 31: 280-283, 1984. PubMed: 6381718

Engel JC, et al. Isolate-dependent differences in the oxidative metabolism of Trypanosoma cruzi epimastigotes. Mol. Biochem. Parasitol. 39: 69-76, 1990. PubMed: 2406595