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Leishmania major (Yakimoff and Schokhor) Bray et al.

PRA-386

Product category
Protists
Product type
Parasitic protozoan
Strain designation
(MHOM/SN/74/SD) Δgp63 1-7 + gp63 1
Type strain
No
Isolation source
Cutaneous leishmaniasis
Geographical isolation
Senegal
Applications
Infectious disease research
Vaccine development
Vector-borne disease research
Product format
Frozen
Storage conditions
-80°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage
Mission Collection Item
This is a Mission Collection Item.

Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed product information

General

Specific applications
Virulence and vaccine studies

Characteristics

Comments
Leishmanolysin-deficient mutant lacking gp63 genes 1-7. Complemented with gp63 gene 1 by transfection. RefJoshi, PB, et al. Targeted gene deletion in Leishmania major identifies leishmanolysin (GP63) as a virulence factor. Mol. Biochem. Parasitol. 120: 33-40, 2002. PubMed: 11849703

Handling information

Medium
Instruction for complete medium
Media: ATCC Medium 2736 should be completed by the addition of 10% HIFBS and 10 μg/mL hemin just prior to use.

Alternate Media: ATCC Medium 1011, ATCC Medium 807 (some strains may not grow equally well in alte
Temperature
20-25°C
Handling procedure
Storage and Culture Initiation
Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen. If liquid nitrogen storage facilities are not available, frozen ampules may be stored at or below -70ºC for approximately one week. Do not under any circumstance store frozen ampules at refrigerator freezer temperatures (generally -20°C). Storage of frozen material at this temperature will result in the death of the culture.
  1. To thaw a frozen ampule, place in a 35°C water bath until thawed (2-3 min). Immerse the ampule just sufficiently to cover the frozen material. Do not agitate the ampule.
  2. Immediately after thawing, aseptically transfer contents to a T-25 tissue culture flask containing 10 mL ATCC Medium 2736. Incubate at 20°C to 25°C with the cap screwed on tightly.
Culture maintenance
  1. Agitate a culture at or near peak density and aseptically transfer 0.1-0.2 mL to a fresh flask of ATCC medium 2736.
  2. Incubate at 20-25°C with the cap screwed on tightly.
  3. Transfer the culture every 7-14 days as described in steps 1-2.  The transfer interval will depend on the quantity of the inoculum and the quality of the medium.  This should be empirically determined by examining the culture on a daily basis until the growth cycle has stabilized.
Cryopreservation
  1. Harvest cells from a culture which is at or near peak density by centrifugation at ~800 x g for 5 min.
  2. Adjust concentration of cells to 2 x 107/mL in fresh medium.  If the concentration is too low, centrifuge at ~800 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired concentration.
  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 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.)  If freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through heat of fusion.  At -40°C, plunge ampules into liquid nitrogen.
  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. Remove the vial from the water bath immediately after thawing.  Aseptically transfer the contents of the ampule into 10 mL of fresh ATCC medium 2736. 
  10. Incubate the flask at 20-25°C with the cap screwed on tightly.
  11. Maintain as described above. 

History

Deposited as
Leishmania major Δgp63 1-7 + gp63 gene 1 NIH SD (MHOM/SN/74/SD)
Depositors
R McMaster
Chain of custody
ATCC <-- Robert McMaster
Year of origin
1973

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Neva FA, et al. Cutaneous leishmaniasis--a case with persistent organisms after treatment in presence of normal immune response. Am. J. Trop. Med. Hyg. 28: 467-71, 1979. PubMed: 222157

McDowell, MA, et al. Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent. Infect. Immun. 70: 3994-4001, 2002. PubMed: 12117904

Joshi, PB, et al. Targeted gene deletion in Leishmania major identifies leishmanolysin (GP63) as a virulence factor. Mol. Biochem. Parasitol. 120: 33-40, 2002. PubMed: 11849703

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