| 产品名称： |
Trypanosoma brucei gambiense Dutton |
| 商品货号： |
TS144190 |
| Deposited As： |
Trypanosoma gambiense Dutton |
| Strain Designations： |
Wellcome Ts |
| 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, Hospital for Tropical Diseases, London, England, 1921 |
| 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： |
Reagents
Tyrodes Salt Solution
NaCl, 8.00 g
KCl, 0.20 g
CaCl2, 0.20 g
MgCl2xa0• H2O, 0.05 g
NaH2PO4xa0• H2O 1.00 g
NaHCO3xa0• H2O, 1.00 g
Glucose, 1.00 g
Glass distilled H2O to 1.00 L
Add ingredients in the sequence listed. Filter-sterilize.
Harvest and Preservation
- Harvest the parasites according to the protocol for maintenancexa0in vivo.
- Spin the cell suspension at approximately 50 x g for 3 min, to remove any cellular debris.
- Transfer the supernatant to a new 15xa0mLxa0plastic 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 107xa0cells/mLxa0with 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% (v/v) DMSO in equal portions.xa0 The final concentration will be 1.0 - 2.0 x 107xa0cells/mLxa0and 5% DMSO.xa0 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.
- Dispense in 0.5xa0mLxa0aliquots to 1.0-2.0xa0mLxa0sterile 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 rat.xa0 Follow the protocol for maintenancexa0in vivo.
|
| Name of Depositor： |
EJ Tobie |
| Chain of Custody： |
ATCC |
| Year of Origin： |
1921 |
| References： |
. . J. Parasitol. 36: 48-54, 1950.
Southworth GG, Read CP. Carbohydrate transport in Trypanosoma gambiense. J. Protozool. 16: 720-723, 1969. PubMed: 5362387
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