Leishmania mexicana (Biagi) Garnham (ATCC® 30880)

Organism: Leishmania mexicana (Biagi) Garnham  /  Depositor: L Hendricks

Deposited As Leishmania braziliensis peruviana
Strain Designations L-677 [UTA, p-Ls 286]
Application
Vector borne research
Biosafety Level 2
Isolation Human, Lima, Peru, 1973
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

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

Medium Medium 1011: Diphasic blood agar medium
Medium 1012: Diphasic blood agar medium
Medium 807: Brain heart infusion blood agar
Medium 431: Trypanosome medium
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 Locke’s solution.  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 more than 15 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 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.)  
  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 thawed.
  9. Immediately after thawing, do not leave in the water bath, 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 L Hendricks
Chain of Custody
ATCC <-- L Hendricks <-- R Zeludon <-- Z Burstain
Year of Origin 1973