To ATCC Valued Customers,

ATCC stands ready to support our customers’ needs during the coronavirus pandemic. Our first job is to listen to and observe what our customers need, and meet those needs with quality products and services. While we are not currently experiencing delays due to this pandemic, we expect that we could see them as the situation evolves. If you experience any issues, please contact ATCC Customer Service at sales@atcc.org. For Technical questions please contact tech@atcc.org. Thank you for your understanding, patience and flexibility as ATCC does everything it can to help reduce the impact of the coronavirus pandemic to our valued customers.
X

Acanthamoeba castellanii (Douglas) Page (ATCC® 30868)

Strain Designations: CCAP 1501/2g  /  Depositor: CCAP  /  Biosafety Level: 2

Permits and Restrictions

View Permits

Strain Designations CCAP 1501/2g
Application
Characterization of plasminogen activator
Contact lens disinfectants against cysts
In vitro characterization of cytopathic effect
Successful immunization against keratitis
characterization of Acanthamoeba polyphaga
Biosafety Level 2

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Isolation
human cornea, Cambridge, England, 1974
Product Format frozen
Type Strain no
Comments
In vivo and in vitro collagenolytic activity
case report
Chemotatic response of macrophages
Susceptibility of corneas from various animal species
Survival in contact lens rinse solution
In vitro intercellular adherence
Characterization of plasminogen activator
mitochondrial DNA fingerprinting
Amebostomes
Antibody dependent neutrophil-mediated killing
Contact lens disinfectants against cysts
In vitro characterization of cytopathic effect
Effect on apoptosis of tumor cells
Successful immunization against keratitis
characterization of Acanthamoeba polyphaga
An axenic version of this strain is available as ATCC 50514
Medium ATCC® Medium 997: Fresh water ameba medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with Escherichia coli
Protocol: ATCCNO: 30011 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in the center of an agar plate of ATCC medium 997. Add the liquid remaining in the vial to the plate and spread it evenly over the surface of the plate. Incubate the plate at 25C. Trophozoites (amebae) should be evident within 2-3 days.
Subcultivation
Protocol: ATCCNO: 30011 SPEC: This strain is distributed as a dried preparation. See the general procedures for opening a dried vial. Aseptically add 1 ml of sterile distilled water to the inner shell vial, remove the filter paper aseptically with a pair of forceps, and place it in the center of an agar plate of ATCC medium 997. Add the liquid remaining in the vial to the plate and spread it evenly over the surface of the plate. Incubate the plate at 25C. Trophozoites (amebae) should be evident within 2-3 days.
Cryopreservation
Cryoprotective Solution

DMSO                                                                                    1.5 ml

Dryl's solution (or similar)                                                  8.5 ml

1.     Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.

2.    Harvest cells from a culture which is at or near peak density by adding 5 ml ATCC medium 5080 (Dryl's solution) and washing cells into suspension.  Rub the surface of the plate with a spread bar to detach adhering trophozoites.

3.     Adjust the concentration of cells to at least 2 x 106/ml in fresh medium.

4.     Mix the cell preparation and the cryoprotective solution in equal portions.

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. 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.     Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8.     To establish a culture from the frozen state place the vial in a 35°C water bath.  Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.   Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.  Distribute the material evenly over the plate using a spread bar.

9.     Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.

Follow the protocol for maintenance of culture.

Mycoplasma Unknown
Name of Depositor CCAP
Special Collection NCRR Contract
Chain of Custody
ATCC <<--CCAP<<--J. Nagington
Year of Origin 1974
References

Silvany RE, et al. The effect of currently available contact lens disinfection systems on Acanthamoeba castellanii and Acanthamoeba polyphaga. Ophthalmology 97: 286-290, 1990. PubMed: 2336265

Naginton J, et al. Amoebic infection of the eye. Lancet 2: 1537-1540, 1974. PubMed: 4140981

Stewart GL, et al. Chemotactic response of macrophages to Acanthamoeba castellanii antigen and antibody-dependent macrophage-mediated killing of the parasite. J. Parasitol. 78: 849-855, 1992. PubMed: 1403427

Nauheim RC, et al. Survival of Acanthamoeba in contact lens rinse solutions. Cornea 9: 290-293, 1990. PubMed: 2127739

Niederkorn JY, et al. Susceptibility of corneas from various animal species to in vitro binding and invasion by Acanthamoeba castellanii [corrected] [published erratum appears in Invest. Ophthalmol. Vis. Sci. 33: 2577, 1992]. Invest. Ophthalmol. Vis. Sci. 33: 104-112, 1992. PubMed: 1730531

He YG, et al. In vivo and in vitro collagenolytic activity of Acanthamoeba castellanii. Invest. Ophthalmol. Vis. Sci. 31: 2235-2240, 1990. PubMed: 2173683

Silvany RE, et al. Effect of contact lens preservatives on Acanthamoeba. Ophthalmology 98: 854-857, 1991. PubMed: 1866136

Ubelaker JE, et al. In vitro intercellular adherence of Acanthamoeba castellanii: A scanning and transmission electron microscopy study. Cornea 10: 299-304, 1991. PubMed: 1889215

Mitra MM, et al. Characterization of a plasminogen activator produced by Acanthamoeba castellanii. Mol. Biochem. Parasitol. 73: 157-164, 1995. PubMed: 8577323

Gautom RK, et al. Mitochondrial DNA fingerprinting of Acanthamoeba spp. isolated from clinical and environmental sources. J. Clin. Microbiol. 32: 1070-1073, 1994. PubMed: 7913095

Ubelaker JE, et al. Amebostomes on the amoeba Acanthamoeba castellanii (Acanthamoebidae: Amoebidae). Trans. Am. Microsc. Soc. 113: 211-215, 1994.

Daggett PM, et al. Distribution and possible interrelationships of pathogenic and nonpathogenic Acanthamoeba from aquatic environments. Microb. Ecol. 8: 371-386, 1982.

Stewart GL, et al. Antibody-dependent neutrophil-mediated killing of Acanthamoeba castellanii. Int. J. Parasitol. 24: 739-742, 1994. PubMed: 7928077

Kilvington S, et al. Effect of contact lens disinfectants against Acanthamoeba cysts. Rev. Infect. Dis. 13 suppl.5: S414-S415, 1991. PubMed: 2047678

Taylor WM, et al. In vitro characterization of Acanthamoeba castellanii cytopathic effect. J. Parasitol. 81: 603-609, 1995. PubMed: 7623204

Alizadeh H, et al. Apoptosis as a mechanism of cytolysis of tumor cells by a pathogenic free-Living amoeba. Infect. Immun. 62: 1298-1303, 1994. PubMed: 8132336

Alizadeh H, et al. Successful immunization against Acanthamoeba keratitis in a pig model. Cornea 14: 180-186, 1995. PubMed: 7743802

Fritsche TR, et al. Occurrence of bacterial endosymbionts in Acanthamoeba spp. isolated from corneal and environmental specimens and contact lenses. J. Clin. Microbiol. 31: 1122-1126, 1993. PubMed: 8501212

Alizadeh H, et al. In vitro amoebicidal activity of propamidine and pentamidine isethionate against Acanthamoeba species and toxicity to corneal tissues. Cornea 16: 94-100, 1997. PubMed: 8985640

Niszl IA, et al. Cytopathogenicity of clinical and environmental Acanthamoeba isolates for two mammalian cell lines. J. Parasitol. 84: 961-967, 1998. PubMed: 9794638

Hurt M, et al. Exacerbation of Acanthamoeba keratitis in animals treated with anti-macrophage inflammatory protein 2 or antineutrophil antibodies. Infect. Immun. 69: 2988-2995, 2001. PubMed: 11292716

Alizadeh H, et al. Tear IgA and serum IgG antibodies against Acanthamoeba in patients with Acanthamoeba keratitis. Cornea 20: 622-627, 2001. PubMed: 11473164

Alves JMP, et al. Random amplified polymorphic DNA probes as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba. Braz. J. Med. Biol. Res. 33: 19-26, 2000.

Aksozek A, et al. Resistance of Acanthamoeba castellanii cysts to physical, chemical, and radiological conditions. J. Parasitol. 88: 621-623, 2002. PubMed: 12099437

Kennett MJ, et al. Acanthamoeba castellanii: characterization of an adhesin molecule. Exp. Parasitol. 92: 161-169, 1999. PubMed: 10403757

Wang X, Ahearn DG. Effect of bacteria on survival and growth of Acanthamoeba castellanii. Curr. Microbiol. 34: 212-215, 1997. PubMed: 9058539

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