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
Bone marrow of 9-year-old Indian male, Bihar, India, 1980
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
Comments
WHO reference strain. Promastigotes.
Selective killing of amastigotes
Medium
ATCC® Medium 807: Brain heart infusion blood agar
ATCC® Medium 1011: Diphasic blood agar medium
ATCC® Medium 1012: Diphasic blood agar medium
ATCC® Medium 431: Trypanosome medium
Growth Conditions
Temperature: 25°C
Culture System: Axenic
Cryopreservation
Harvest and Preservation
Harvest cells from a culture which is at or near peak density by centrifugation at 1,300 g for 5 min.
Adjust concentration of cells to 2 x 107/mL in fresh medium.
While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium (broth). The DMSO solution when first prepared will warm up due to chemical heat. The solution should be allowed to return to room temperature prior to use.
Mix the cell preparation and the DMSO solution in equal portions. The final concentration will be 107 cells/mL and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no more than 15 min.
Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
Place the ampules 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, do not leave in the water bath, aseptically transfer the contents of the ampule into a fresh tube of ATCC medium 807.
Incubate vertically at 25°C with the cap screwed on tightly.
Maintain as described above.
Name of Depositor
WRAIR
Chain of Custody
ATCC <-- WRAIR <-- W. Peters <-- L. Srivistava
Year of Origin
1980
References
Xu ZB, et al. The characterization by isoenzyme electrophoresis of Leishmania isolated in the People's Republic of China. Trans. R. Soc. Trop. Med. Hyg. 78: 689-693, 1984. PubMed: 6506159
Muyombwe A, et al. Selective killing of Leishmania amastigotes expressing a thymidine kinase suicide gene. Exp. Parasitol. 85: 35-42, 1997. PubMed: 9024200
