| 产品名称： |
Trypanosoma brucei rhodesiense Stephens and Fantham |
| 商品货号： |
TS161435 |
| Deposited As： |
Trypanosoma rhodesiense Stephens and Fantham |
| Strain Designations： |
Uganda 105A |
| Application： |
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. |
| Isolation： |
Human, Uganda, 1959 |
| 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 |
| Type Strain： |
no |
| Growth Conditions： |
Culture System: in vivo, laboratory rat |
| Cryopreservation： |
Harvest and Preservation
- Harvest the parasites according to the protocol for maintenance in vivo.
- Spin the cell suspension at approximately 50 x g for 3 min, to remove the cellular debris.
- Transfer the supernatant to a new 15 mL plastic centrifuge tube.xa0 Centrifuge at 1300 x g for 10 min.
- Pool the cell pellets and adjust the concentration to 2.0 - 4.0 x 107 cells/mL with a fresh solution of Tyrodes Salt Solution.xa0*If the concentration is too low centrifuge at 1300 x g for 10 min and resuspend in the volume of Tyrodes Salt Solution required to yield the desired concentration.
- Mix the cell preparation and 10% DMSO (v/v) Tyrodes Salt Solution in equal portions.xa0 The final concentration will be 1.0 - 2.0 x 107 cells/mL and 5% DMSO in Tyrodes Salt Solution.xa0 The time from the mixing of the cell preparation and the cryoprotective solution before the freezing process begins should be no less than 15 min. and no more than 30 min.
- Dispense in 0.5 mL aliquots to 1.0-2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
- Place the vials in a controlled rate freezing unit.xa0 From room temperature cool 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 the heat of fusion.xa0 At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.xa0 Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximatelyxa0-1°C/min.) xa0
- Store in either the vapor or liquid phase of a nitrogen refrigerator.
- 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.xa0 Do not agitate the ampule.xa0 Do not leave ampule in water bath after thawed.
- Immediately after thawing, aseptically remove the contents of the ampule with a syringe and inoculate an uninfected mouse.xa0 Follow the protocol for maintenance in vivo.
|
| Name of Depositor： |
RG Yaeger |
| Chain of Custody： |
ATCC |
| Year of Origin： |
1959 |
| References： |
. . Exp. Parasitol. 13: 374-385, 1963.
|
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