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Leishmania donovani (Laveran and Mesnil) Ross

PRA-413

Product category
Protists
Product type
Parasitic protozoan
Strain designation
AG83 [MHOM/IN/1983/AG83]
Isolation source
Bone marrow aspirate from patient with Kala-azar
Geographical isolation
India
Applications
Infectious disease research
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

Characteristics

Comments
Strain is also known as MHOM/IN/1983/AG83

Handling information

Medium
Instruction for complete medium
Media: ATCC Medium 2736

Alternate Media: ATCC Medium 431 Trypanosome medium, ATCC Medium 1011 Diphasic blood agar medium (30% rabbit blood), ATCC Medium 1012 Diphasic blood agar medium (10% rabbit blood), ATCC
Temperature
20-25°C
Culture system
Axenic
Handling procedure
Storage and Culture Initation
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 ampoules may be stored at or below -70°C 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 sufficient 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 10.0 mL ATCC medium 2736. Incubate at 20-25°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 that is at or near peak density by centrifugation at ~800 x g for 5 min. Pool the cell pellets into a single tube.
  2. Adjust the concentration of cells to 2.0 x 107/mL.  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. Prepare a 10% (v/v) sterile DMSO solution in fresh medium as follows:  Add the required volume of DMSO to a glass screw-capped test tube and place on ice.  Allow the DMSO to solidify.  Add the required volume of refrigerated medium.  Dissolve the DMSO by inverting several times.  If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
  4. Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 107 and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution to the start of the freezing process should be no less than 15 min and no longer than 60 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 vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. 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. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen refrigerator.
  8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial enough to cover only the frozen material. Do not agitate the vial.
  9. Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate into 10.0 mL of fresh ATCC medium 2736.
  10. Incubate the tube at 20-25°C with the cap screwed on tightly.

History

Deposited as
Leishmania donovani
Depositors
N Ali
Chain of custody
ATCC <-- N Ali
Type of isolate
Human
Year of origin
1983

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

Mukhopadhyay S, et al. Immunoprophylaxis and immunotherapy against experimental visceral leishmaniasis. Vaccine 17(3): 291-300, 1999. PubMed: 9987166

Mukhopadhyay S, et al. Use of an attenuated leishmanial parasite as an immunoprophylactic and immunotherapeutic agent against murine visceral leishmaniasis. Clin. Diagn. Lab Immunol. 7(2): 233-240, 2000. PubMed: 10702498

Pucadyil TJ, et al. Cholesterol is required for Leishmania donovani infection: implications in leishmaniasis. Mol. Biochem. Parasitol. 133(2): 145-152, 2004. PubMed: 14698427

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