Leishmania major (Yakimoff and Schokhor) Bray et al. (ATCC® 30012)

Organism: Leishmania major (Yakimoff and Schokhor) Bray et al.  /  Depositor: LS Diamond

Deposited As Leishmania tropica (Wright) Luhe
Strain Designations No designation
Application
Vector borne research
Biosafety Level 2
Isolation Human, Teheran, Iran, 1949
Product Format frozen
Storage Conditions Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Comments
Promastigotes
lipids

Cyclopropane fatty acid

Sequencing of ITS1, 5.8S rRNA, and ITS2 regions performed at ATCC shows 99% identity to Leishmania major.

 

Medium Medium 1011: Diphasic blood agar medium
Medium 1012: Diphasic blood agar medium
Medium 807: Brain heart infusion blood agar
Growth Conditions
Temperature: 25°C
Culture System: Axenic
Cryopreservation Harvest and Preservation
  1. Harvest cells from a culture which is at or near peak density by centrifugation at 1,300 g for 5 min.
  2. Adjust concentration of cells to 2 x 107/mL in fresh medium.
  3. 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.
  4. 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 less than 15 min and no more than 30 min.
  5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
  6. 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.)  
  7. Store in either the vapor or liquid phase of a nitrogen refrigerator.
  8. 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 it is thawed.
  9. Immediately after thawing, do not leave in the water bath; rather, aseptically transfer the contents of the ampule into a fresh tube of ATCC medium 1011. 
  10. Incubate vertically at 25°C with the cap screwed on tightly.
  11. Maintain as described above. 
Name of Depositor LS Diamond
Chain of Custody
ATCC <-- LS Diamond<-- E Tobie <-- Dr. Ansary <-- U.S. Army Med. Lab
Year of Origin 1949
References

Beach DH, et al. Lipids of Leishmania promastigotes. J. Parasitol. 65: 201-216, 1979. PubMed: 448607

Fish WR, et al. The cyclopropane fatty acid of trypanosomatids. Mol. Biochem. Parasitol. 3: 103-115, 1981. PubMed: 7254247