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Product Name:
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Trypanosoma cruzi
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Manufacturer:
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BEI Resources
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Taxonomy:
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Protozoa Classification: Trypanosomatidae, Trypanosoma
Species: Trypanosoma cruzi
Strain: CL-Luc::Neon/Cas9
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Additional Information:
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The protozoan parasite T. cruzi is the causative agent of Chagas’ disease, a debilitating vectorborne disease endemic in North, Central and South America.3 In North America, T. cruzi has been identified through climactic and vector-based data as a potential emerging health risk to humans in the southern United States, where the two most commonly reported reservoirs in North America are the raccoon and the Virginia opossum.4,5 The parasite has a complex life cycle and is transmitted by hematophagous triatomine reduviid bugs to wildlife and exotic mammal species, domestic dogs, and humans.4,5 Dogs are considered a reservoir in the domestic transmission cycle of T. cruzi in endemic areas.4,6
T. cruzi is currently classified into six discrete typing units (TcI, TcII, TcIII, TcIV, TcV and TcVI), which are identifiable by common molecular markers and represent different eco-epidemiological features, pathogenicity and geographical distribution.1,7 The parent strain (NR-49381) corresponds to discrete typing unit (DTU) T. cruzi VI (TcVI) based on sequence analysis of the putative C-5 sterol desaturase gene, TcSC5D.7 TcVI is considered a hybrid DTU containing both TcII and TcIII genomes, and is most associated with domestic infection cycles of Chagas’ disease in southern and central South America.7
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Material Provided:
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Each vial of NR-59633 contains approximately 0.5 mL of culture in cryopreservative [5% dimethylsulfoxide (DMSO)]. Please see Appendix I for cryopreservation instructions.
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Packing/Storage:
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NR-59633 was packaged aseptically in cryovials and is provided frozen on dry ice. The product should be stored at -130°C or colder, preferably in the vapor phase of a liquid nitrogen freezer. If liquid nitrogen storage facilities are not available, frozen cryovials may be stored at -70°C or colder for approximately one week.
Note: Do not under any circumstances store vials at temperatures warmer than -70°C. Storage under these conditions will result in the death of the culture.
To ensure the highest level of viability, the culture should be initiated immediately upon receipt. Any warming of the product during shipping and transfer must be avoided, as this will adversely affect the viability of the product. For transfer between freezers and for shipping, the product may be placed on dry ice for brief periods, although use of a portable liquid nitrogen carrier is preferred. Please read the following recommendations prior to using this material.
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Growth Conditions:
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Media:
Liver Infusion Tryptose (LIT) medium (Appendix II) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (HIFBS) and 10 µg/mL hemin
Incubation:
Temperature: 37°C
Atmosphere: Aerobic
Propagation:
1. To establish a culture from the frozen state, place a vial in a 35°C to 37°C water bath. Thawing time is approximately 2 to 3 minutes. Do not agitate the vial. Do not leave the vial in the water bath after it is thawed.
2. Immediately after thawing, aseptically transfer the contents to a tissue culture flask containing 10 mL of LIT medium. Incubate at 25°C with the cap screwed on tightly.
3. Observe the culture daily under an inverted microscope for the presence of epimastigote forms of the parasite. Subculture when the culture has reached peak density.
Maintenance:
1. Agitate a culture at or near peak density and aseptically transfer 0.5 mL to 1.0 mL into a new tissue culture flask with fresh growth medium.
2. Incubate the culture at 25°C with the cap screwed on tightly and examine daily under an inverted microscope.
3. Transfer every 3 to 7 days, as needed. Note that the transfer interval should be determined empirically as it is dependent on the quantity of the inoculum.
Please refer to Appendix I for cryopreservation instructions.
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Disclaimers:
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You are authorized to use this product for research use only. It is not intended for human use. Use of this product is subject to the terms and conditions of the BEI Resources Material Transfer Agreement (MTA). The MTA is available on our Web site at www.beiresources.org. While BEI Resources uses reasonable efforts to include accurate and up-to-date information on this product sheet, neither ATCC® nor the U.S. Government makes any warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. Neither ATCC® nor the U.S. Government warrants that such information has been confirmed to be accurate. This product is sent with the condition that you are responsible for its safe storage, handling, use and disposal. ATCC® and the U.S. Government are not liable for any damages or injuries arising from receipt and/ or use of this product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, the U.S. Government, ATCC®, their suppliers and contributors to BEI Resources are not liable for damages arising from the misidentification or misrepresentation of products. |
References:
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1. Kelly, R. M., Personal Communication.
2. Costa, F.C., et al. “Expanding the Toolbox for Trypanosoma cruzi: A Parasite Line Incorporating a Bioluminescence-fluorescence Dual Reporter and Streamlined CRISPR/Cas9 Functionality for Rapid In Vivo Localisation and Phenotyping." PLoS Negl Trop Dis. 12 (2018): e0006388. PubMed: 29608569.
3. Bern, C. “Chagas’ Disease.” N. Engl. J. Med. 373 (2015): 456-466. PubMed: 26222561.
4. Patel, J. M., et al. “Isolation, Mouse Pathogenicity, and Genotyping of Trypanosoma cruzi from an English Cocker Spaniel from Virginia, USA.” Vet. Parasitol. 187 (2012): 394-398. PubMed: 22341614.
5. Brown, E. L., et al. “Seroprevalence of Trypanosoma cruzi among Eleven Potential Reservoir Species from Six States across the Southern United States.” Vector Borne Zoonotic Dis. 10 (2010): 757-763. PubMed: 20020815.
6. Estrada-Franco, J. G., et al. “Human Trypanosoma cruzi Infection and Seropositivity in Dogs, Mexico.” Emerg. Infect. Dis. 12 (2006): 624-630. PubMed: 16704811.
7. Zingales, B., et al. “The Revised Trypansoma cruzi Subspecific Nomenclature: Rationale, Epidemiological Relevance and Research Applications.” Infect. Genet. Evol. 12 (2012): 240-253. PubMed: 22226704.
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Citation:
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Acknowledgment for publications should read “The following reagent was obtained through BEI Resources, NIAID, NIH: Trypanosoma cruzi, Strain CL-Luc::Neon/Cas9, NR-59633.”
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Biosafety Level:
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2
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories (BMBL). Current Edition. Washington, DC: U.S. Government Printing Office.
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