Crithidia oncopelti (Noguchi and Tilden) Hanson and McGhee (ATCC® 12982)

Depositor: HN Guttman  /  Biosafety Level: 1

Permits and Restrictions

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Biosafety Level 1
Isolation
Oncopeltus fasciatus (?), 1926
Product Format frozen
Type Strain no
Comments
Composition of paraflagellar rod
monophyly of endosymbiont-containing trypanosomatids
Ultrastructure of symbiotic bacteria
Riboprinting and taxonomy
Effects of methylglyoxal bis (guanylhydrazone) on growth
Cyclopropane fatty acid
Reduced growth of symbiotes free cells
Symbiote-free hemoflagellates, liver factor requirement and serologic identity
Medium Medium 483: Guttman's IIB 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 1034. 

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 1034 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 1034 in a 16 x 125 mm screw-capped test tube. Incubate upright at 25°C with caps screwed on tightly.

Name of Depositor HN Guttman
Chain of Custody
ATCC <<--HN Guttman<<--M. Lwoff <<--- C.M. Wenyon <<--- H. Noguchi
Year of Origin 1926
References

Chang KP. Ultrastructure of symbiotic bacteria in normal and antibiotic-treated Blastocrithidia culicis and Crithidia oncopelti. J. Protozool. 21: 699-707, 1974. PubMed: 4217371

Chang KP, et al. Effects of methylglyoxal bis(ganylhydrazone) on trypanosomatid flagellates: inhibition of growth and nucleoside incorporation in Trypanosoma brucei. J. Protozool. 25: 145-149, 1978. PubMed: 660567

. . Exp. Parasitol. 14: 129-142, 1963.

Schlaeppi K, et al. The major component of the paraflagellar rod of Trypanosoma brucei is a helical protein that is encoded by two identical, tandemly linked genes. J. Cell Biol. 109: 1695-1709, 1989. PubMed: 2793936

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

Goncanlves De Lima VM, et al. Comparison of six isoenzymes from 10 species of Crithidia. J. Protozool. 29: 397-401, 1982.

Fish WR, et al. The cyclopropane fatty acid of trypanosomatids. Mol. Biochem. Parasitol. 3: 103-115, 1981. PubMed: 7254247

Chang KP. Reduced growth of Blastocrithidia culicis and Crithidia oncopelti freed of intracellular symbiotes by chloramphenicol. J. Protozool. 22: 271-276, 1975. PubMed: 807721

Chang KP. Symbiote-free hemoflagellates, Blastocrithidia culicis and Crithidia oncopelti: their liver factor requirement and serologic identity. J. Protozool. 23: 241-244, 1976. PubMed: 933081

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

Hollar J, et al. Monophyly of endosymbiont containing trypanosomatids: Phylogeny versus taxonomy. J. Eukaryot. Microbiol. 45: 293-297, 1998. PubMed: 9627990

Cross References

Nucleotide (GenBank) : AF060882 Crithidia oncopelti kinetoplast NADH dehydrogenase subunit 8 (ND8) and Cyb gRNA genes, complete cds.

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