Trypanosoma cruzi Chagas
PRA-333 ™
- Product category
-
Protists
- Product type
-
Parasitic protozoan
- Classification
- KINGDOM: Protozoa
- Strain designation
- VL2067 LacZ clone A8
- Type strain
- No
- Isolation source
- Transgenic clone derived from VL2067 strain
- Product format
- Frozen
Documentation
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Detailed product information
Characteristics
- Comments
-
Trypomastigotes expressing beta-galactosidase
Handling information
- Host
-
BALB/3T3 clone A31 (ATCC CCL-163)
- Medium
- Temperature
-
35-37°C
- Cryopreservation
-
1. Harvest Trypanosoma cultures when emergent parasites (trypomastigote stage) have reached or are near peak density in the liquid column. Gently invert the Trypanosoma culture flasks to suspend parasites in the liquid medium.
2. Transfer the cell suspension (including parasites) to 15 ml plastic centrifuge tubes. Centrifuge at 1300 x g for 10 min.
3. Remove all but 0.5 ml of the supernatant from each tube, resuspend the cell pellets, and pool them to a single tube.
4. Adjust the parasite concentration to 2.0 - 4.0 x 107 cells/ml with fresh medium or PBS.
NOTE: If the concentration of parasites is too low, centrifuge at 1300 x g for 10 min and resuspend in the volume of fresh medium or PBS required to yield the desired concentration.
5. Prepare a cryoprotective solution containing 10% (v/v) DMSO in fresh medium or PBS.
6. Mix the cell preparation and cryoprotective solution in equal portions. The final concentration will be 1.0 - 2.0 x 107 cells/ml and 5% DMSO. The time from the mixing of the cell preparation and cryoprotective solution to the start of the freezing process should be no less than 15 min and no more than 30 min.
NOTE: To prevent culture contamination, penicillin-streptomycin solution (ATCC® 30-2300) may be added to a final concentration of 50 to 100 I.U./ml penicillin and 50 to 100 µg/ml streptomycin.
7. Dispense in 0.5 ml aliquots to 1.0-2.0 ml sterile plastic screw-capped cryovials.
8. Place cryovials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1° C/min through heat of fusion. 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 frozen ampules in either the vapor or liquid phase of a nitrogen refrigerator.
10. To thaw a frozen ampule, place it in a 35-37°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.
11. Immediately after thawing, aseptically transfer contents to a T-25 tissue culture flask containing a fresh monolayer of ATCC® CCL-163 cells and 10 ml ATCC® 30-2002 with 10% (v/v) HIFBS.
12. Outgas the flask for 10 seconds with a 95% air, 5% CO2 gas mixture.
13. Incubate in a 35-37°C CO2 incubator with the cap screwed on tightly.
History
- Depositors
- FS Buckner
- Year of origin
- 1998
Legal disclaimers
- Intended use
- This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
- Warranty
-
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
- Disclaimers
-
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
Permits & Restrictions
Import Permit for the State of Hawaii
If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.
Frequently Asked Questions
References
Curated Citations
Buckner FS, et al. Potent anti-Trypanosoma cruzi activities of oxidosqualene cyclase inhibitors. Antimicrob. Agents Chemother. 45: 1210-1215, 2001. PubMed: 11257036
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