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Product Name:
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STIB 247
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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 brucei subsp. brucei
Strain: STIB 247
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Additional Information:
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T. brucei is a kinetoplastid protozoan parasite and is the causative agent of African trypanosomiasis, which is transmitted to both humans and livestock through the bite of the tsetse fly.5,6 The flies inject the infective stage (metacyclic trypomastigotes) from their salivary glands into the blood and lymphatic fluid of the host, where they undergo differentiation and enter the central nervous system by evading the host immune system through the use of antigenic variation of their surface glycoprotein coat.5,6
T. brucei is divided into three morphologically-identical subspecies that demonstrate distinct pathogenicities: T. brucei subsp. gambiense and T. brucei subsp. rhodesiense, which cause African sleeping sickness in humans, and the non-human infective T. brucei subsp. brucei.5,7 The production of vaccines against these diseases is difficult, as a significant rise of resistance to trypanocidal drugs has been documented. Animal models, such as mice, are critical to understanding the mechanisms of the disease and aid in the development of new therapeutic drugs.5
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Material Provided:
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Each vial of NR-36197 contains approximately 0.5 mL of trypanosome-infected blood with 20% glycerol, 6.25% Yaeger’s anticoagulant and Trypanosome dilution buffer. Please see the Appendix for cryopreservation instructions and component details.
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Packing/Storage:
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NR-36197 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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In vivo, BALB/c mouse or Fox Chase SCID mouse or equivalent
Inoculation:
1. Thaw a frozen ampule of NR-36197 in a 35°C to 37°C water bath for approximately 2 to 3 minutes.
2. Remove the contents of the ampule using a 1 mL syringe equipped with a needle.
3. Inject the contents of the vial into one mouse by facility-approved methods.
Monitoring parasitemia:
1. Bleed the mouse at 1- to 3-day intervals to monitor parasitemia by microscopic examination using a haemocytometer and 0.85% ammonium chloride as diluent. Parasitemia may also be assessed by microscopic examination of blood films stained with Giemsa solution.
2. Passage the strain when the infection is at or near the first peak of parasitemia (≥ 1 × 107 parasites/mL or ≥ 10 parasites/high power field for Giemsa-stained blood films observed under 100×). This will normally occur after 4 to 5 days of inoculation. Note that the rate of T. brucei subsp. brucei infection may vary with the parasite strain and concentration of inoculum.
Passaging:
1. Anesthetize the infected mouse and collect blood by facility-approved methods. Collect the blood using an anticoagulant such as Yaeger's anticoagulant solution (Appendix I) or EDTA.
2. Perform a parasite count and inject 5 × 105 to 1 × 106 parasites into each of the uninfected mice (approximately 10 mice).
3. Monitor parasitemia as described above and passage as needed.
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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. Geigy, R. and M. Kauffmann. “Sleeping Sickness Survey in the Serengeti Area (Tanzania) 1971. I. Examination of Large Mammals for Trypanosomes.” Acta Trop. 30 (1973): 12-23. PubMed: 4144952.
2. Laboratory of Molecular Parasitology - The Rockefeller University
3. Scott, A. G., et al. “Characterisation of Cloned Lines of Trypanosoma brucei Expressing Stable Resistance to MelCy and Suramin.” Acta Trop. 60 (1996): 251-262. PubMed: 8659324.
4. Jenni, L., et al. “Hybrid Formation between African Trypanosomes during Cyclical Transmission.” Nature. 322 (1986): 173-175. PubMed: 3724860.
5. Antoine-Moussiaux, N., et al. “Contributions of Experimental Mouse Models to the Understanding of African Trypanosomiasis.” Trends Parasitol. 24 (2008): 411-418. PubMed: 18684669.
6. Peacock, L., et al. “Identification of the Meiotic Life Cycle Stage of Trypanosoma brucei in the Tsetse Fly.” Proc. Natl. Acad. Sci. USA 108 (2011): 3671-3676. PubMed: 21321215.
7. Turner, C. M., et al. “Replication, Differentiation, Growth and the Virulence of Trypanosoma brucei Infections.” Parasitology. 111 (1995): 289-300. PubMed: 7567097.
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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 brucei subsp. brucei, Strain STIB 247, NR-36197."
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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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