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
Mouse blood, Greenwich, CT, 2004.
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
Genome Sequenced Strain
Yes
Comments
Genome sequencing strain
Growth Conditions
Culture System:In vivo, SCID mouse
Cryopreservation
Harvest and Preservation
Prepare a 30% (v/v) sterile glycerol solution in Alsever's solution.
Dispense 0.5 mL of anticoagulant solution into a 15 mL test tube. Add to the anticoagulant blood collected by orbital bleeding from SCID mice that had reached or are near peak parasitemia. Invert the tube several times to mix the blood with the anticoagulant.
Mix the heparinized blood with the 30% glycerol solution in a 2:1 ratio. If any clotting has occurred do not use. After mixing, the final concentration of cryoprotectant solution will be 10% (v/v). The mixture should be placed in a 4°C ice bath. The time from the mixing of the cell preparation and glycerol stock solution before the freezing process is begun should be no less than 15 min and no longer than 30 min.
Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryovials. Filled ampules should be placed in a 4°C ice bath. Do not immerse ampules to the level of the vial cap.
Plunge ampules from 4°C into liquid nitrogen. The frozen preparations may be stored in a mechanical freezer until needed, however storage in either the vapor or liquid phase of a nitrogen refrigerator is recommended for the longest viability.
To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min). Immerse the ampule just sufficient to cover the frozen material. Do not agitate the ampule.
Immediately after thawing, aseptically remove the contents of the ampule with a syringe and inoculate an uninfected SCID mouse. Follow the protocol for maintenance in vivo. The course of infection may be longer or shorter than usual depending on recovery of the parasite from the frozen state.
Name of Depositor
C Ben Mamoun
Chain of Custody
ATCC <-- C Ben Mamoun
References
Anderson JF, Magnarelli LA. Babesiosis in Fairfield County, Connecticut. Emerg. Infect. Dis. 10(3): 545-546, 2004. PubMed: 15116708
