| 产品名称： |
Trypanosoma brucei rhodesiense Stephens and Fantham |
| 商品货号： |
TS205030 |
| Deposited As： |
Trypanosoma brucei rhodesiense |
| Strain Designations： |
KETRI 269 |
| 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 clinical isolate, Kitanga, Tanzania, 1960 |
| 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 |
| Comments： |
Produces acute and fatal parasitemia in mice |
| Growth Conditions： |
Culture System: In vivo, Balb/c mouse |
| Cryopreservation： |
Reagents
Trypanosome Dilution Buffer
20 mM Na2HPO4
2 mM NaH2PO4
80 mM NaCl
5 mM KCl
1 mM MgSO4
20 mM glucose
Adjust the pH of the solution to 7.7 and filter sterilize.
Harvest and Preservation
- Prepare a 40% (v/v) sterile glycerol solution in Trypanosome Dilution Buffer (TDB).
- Dispense 0.5 mL of anticoagulant solution into a 15 mL test tube. Add to the anticoagulant blood collected by orbital bleeding from mice that had reached or are near peak parasitemia. Invert the tube several times to mix the blood with the anticoagulant.xa0
- In a separate test tube, add the heparinized blood dropwise to the 40% glycerol solution. Note that blood should be mixed with glycerol solution in a 1:1 ratio to obtain a final concentration of cryoprotectant of 20% (v/v).xa0 Mix slowly by inversion and place the tube on ice.xa0 The freezing process should start 15 to 30 minutes following the addition of the heparinized blood to the cryoprotectant solution.
- Dispense 0.5 mL aliquots of blood suspension into 1.0 - 2.0 mL sterile plastic screw-capped cryovials.xa0 Place the vials in a controlled rate freezing unit.xa0 From room temperature cool the vials at -1°C/min to -40°C.xa0 If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through this phase.xa0 At -40°C, plunge vials into liquid nitrogen. Alternatively, place the vials in a Nalgene 1oC freezing container.xa0 Place the container at -80°C for 1.5 to 2 hours and then plunge vials into liquid nitrogen.
- To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min).xa0 Immerse the ampule just sufficient to cover the frozen material.xa0 Do not agitate the ampule.
- Immediately after thawing, aseptically remove the contents of the ampule with a syringe and inoculate a Balb/c mouse.xa0 Follow the protocol for maintenance of the culture above. xa0
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| Name of Depositor： |
CJ Bacchi |
| References： |
Bacchi CJ, et al. Differential susceptibility to DL-alpha-difluoromethylornithine in clinical isolates of Trypanosoma brucei rhodesiense. Antimicrob Agents Chemother 34(6): 1183-1188, 1990. PubMed: 2118325
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