Trypanosoma brucei rhodesiense Stephens and Fantham
商品貨號(hào)
B215489
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. 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 Tyrode's Salt Solution. *If the concentration is too low centrifuge at 1300 x g for 10 min and resuspend in the volume of Tyrode's Salt Solution required to yield the desired concentration.
Mix the cell preparation and 10% DMSO (v/v) Tyrode's Salt Solution in equal portions. The final concentration will be 1.0 - 2.0 x 107 cells/mL and 5% DMSO in Tyrode's Salt Solution. 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. From room temperature cool at -1°C/min to -40°C. If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion. At -40°C plunge 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.)
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. Do not agitate the ampule. 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. Follow the protocol for maintenance in vivo.
Name of Depositor
RG Yaeger
Chain of Custody
ATCC <-- RG Yaeger <-- F. Goble LSH and TM 105A <-- . . . <-- D.H.H. Robertson EATRO 105
