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Crithidia fasciculata Leger

50083

Product category
Protists
Product type
Parasitic protozoan
Strain designation
ReF-1:PRR
Type strain
No
Isolation source
Cog-wheel bug, Arilus cristatus, from Dorchester Co.
Geographical isolation
United States; Maryland
Product format
Frozen
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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
susceptibility testing iodoquinol
susceptibility testing metronidazole
susceptibility testing paromomycin aminosidine, catenulin
susceptibility testing tetracycline

Characteristics

Comments
Used in monoxenic culture and as an intermediate step in axenization of Entamoeba.
Improved method for the monoxenic cultivation
Riboprinting and taxonomy
Use as food in monoxenic culture
Axenic cultivation
Multiple distinct site-specific elements in miniexon arrays

Handling information

Medium
Instruction for complete medium
Media: ATCC Medium 355.

Alternate Media: ATCC Medium 1034 can also be used for cultivation and is available in a freeze-dried format from ATCC.
Temperature
25°C
Culture system
Axenic
Culture maintenance
1.   When the culture is at or near peak density, vigorously agitate the culture.

2. Transfer approximately 0.10 ml to a fresh tube containing 5 ml of fresh ATCC medium 355. 

3.   Incubate upright at 25°C with caps screwed on tightly.

4.   Transfer every 14 days.

Cryopreservation

1.   Prepare a 10% (v/v) sterile DMSO solution in fresh ATCC Medium 355. 

2.   Transfer a culture at peak density to centrifuge tubes and centrifuge at 525 x g for 5 minutes.

3.   Remove the supernatant and resuspend the cells in ATCC medium 355 to a concentration of 2 x 106 to 2 x 107 cells/ml.

4.   Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/ml and 5% (v/v) DMSO.

5.   Distribute the cell suspension in 0.5 ml aliquots into 1.0-2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).  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 longer than 30 min.

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. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.

8.   To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial.

9.   Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5 ml of fresh ATCC medium 355 in a 16 x 125 mm screw-capped test tube. Incubate upright at 25°C with caps screwed on tightly.

History

Deposited as
Crithidia fasciculata Leger
Depositors
LS Diamond
Type of isolate
Arthropod
Year of origin
1958

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

Gannon JT, Linke HA. Growth studies on xenic cultures of Entamoeba gingivalis using established media. Int. J. Parasitol. 19: 835-838, 1989. PubMed: 2635159

Diamond LS. Improved method for the monoxenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae with trypanosomatids. J. Parasitol. 54: 715-719, 1968. PubMed: 4319344

Raether W, et al. Adaption of amoebae-Crithidia-cultures (Entamoeba histolytica) to axenic conditions of cultivation in TP-S-1-medium of Diamond 1968 (author's transl). Z. Parasitenkd. 42: 279-291, 1973. PubMed: 4360330

Clark CG. Riboprinting: A tool for the study of genetic diversity in microorganisms. J. Eukaryot. Microbiol. 44: 277-283, 1997. PubMed: 9225441

Clark CG. Axenic Cultivation of Entamoeba dispar Brumpt 1925, Entamoeba insolita Geiman and Wichterman 1937 and Entamoeba ranarum Grassi 1879. J. Eukaryot. Microbiol. 42: 590-593, 1995. PubMed: 7581333

View All Curated Citations for this Product

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