Crithidia fasciculata Leger (ATCC® 50083)

Organism: Crithidia fasciculata Leger  /  Depositor: LS Diamond

Permits and Restrictions

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Strain Designations ReF-1:PRR
Application
susceptibility testing iodoquinol
susceptibility testing metronidazole
susceptibility testing paromomycin aminosidine, catenulin
susceptibility testing tetracycline
Biosafety Level 1
Isolation
cog-wheel bug, Arilus cristatus, Dorchester Co., MD, 1958
Product Format frozen
Type Strain no
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
Medium ATCC® Medium 1373: TTYSH medium
ATCC® Medium 1373: TTYSH medium
ATCC® Medium 355: Crithidia medium
Growth Conditions
Temperature: 25.0°C
Duration: axenic
Protocol: ATCCNO: 11745 SPEC: See general instructions for thawing and storage of frozen material before proceeding. Add thawed contents to a single 16 x 125 mm glass screw-capped test tube of the appropriate medium. Incubate the culture vertically with the cap screwed on tightly. It is essential to establish cultures initially in small volumes. Once established, the culture can be scaled up to larger volumes. Vigorously agitate the culture and aseptically transfer 0.1 ml of culture to a fresh tube of medium weekly.
Subcultivation
Protocol: ATCCNO: 11745 SPEC: See general instructions for thawing and storage of frozen material before proceeding. Add thawed contents to a single 16 x 125 mm glass screw-capped test tube of the appropriate medium. Incubate the culture vertically with the cap screwed on tightly. It is essential to establish cultures initially in small volumes. Once established, the culture can be scaled up to larger volumes. Vigorously agitate the culture and aseptically transfer 0.1 ml of culture to a fresh tube of medium weekly.
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.

Name of Depositor LS Diamond
Year of Origin 1958
References

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

Chan FT, et al. Susceptibility testing of Dientamoeba fragilis ATCC 30948 with iodoquinol, paromomycin, tetracycline, and metronidazole. Antimicrob. Agents Chemother. 38: 1157-1160, 1994. PubMed: 8067755

Teng SC, et al. A new non-LTR retrotransposon provides evidence for multiple distinct site-specific elements in Crithidia fasciculata miniexon arrays. Nucleic Acids Res. 23: 2929-2936, 1995. PubMed: 7659515

Clark CG, et al. Entamoeba histolytica: is conversion of "nonpathogenic" amebae to the "pathogenic" form a real phenomenon?. Exp. Parasitol. 74: 307-314, 1992. PubMed: 1582483

Cho J, Eichinger D. Crithidia fasciculata induces encystation of Entamoeba invadens in a galactose-dependent manner. J. Parasitol. 84: 705-710, 1998. PubMed: 9714198

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation