Acanthamoeba castellanii (Douglas) Page (ATCC® 30010)

Organism: Acanthamoeba castellanii (Douglas) Page  /  Depositor: RJ Neff

Deposited As Acanthamoeba sp.
Strain Designations Neff
Application Control strain
Editing of tRNAs
Oxidative metabolism associated with phagocytosis
Inhibition by Pseudomonas aeruginosa
Quantitative bacterial plaque assay for enumeration
Chlorhexidine tablet system for disinfection
Internalisation of bacterial substrate as factors in growth rate
Production of monoclonal antibodies
Biosafety Level 1
Isolation Soil, Pacific Grove, CA, 1957
Product Format frozen
Storage Conditions Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Comments
Very similar to A. terricola ATCC 30134 on the basis of starch gel electrophoretic analysis of enzymes.
Two genetic markers that distinguish pathogenic and nonpathogenic strains
resistant to complement lysis
Editing of tRNAs
Nucleotide sequences of 5S and 5.8S RNAs
Isoenzyme electrophoresis
Axenic cultivation
Oxidative metabolism associated with phagocytosis
Biochemical and molecular characterization
characterization of Acanthamoeba polyphaga
inhibition by Pseudomonas aeruginosa
quantitative bacterial plaque assay for enumeration
Chlorhexidine tablet system for disinfection
Interaction of Legionella pneumophila
Internalisation of bacterial substrate as factors in growth rate
Production of monoclonal antibodies
Occurrence in natural thermal waters in Mexico
review
phylogeny
Medium ATCC® Medium 712: PYG w/ Additives
Growth Conditions
Temperature: 25°C
Culture System: Axenic
Cryopreservation

Harvest and Preservation

  1. To achieve the best results, set up cultures with several different inocula (e.g. 0.25 mL, 0.5 mL, 1.0 mL). Harvest cultures and pool when the culture that received the lowest inoculum is at or near peak density.
  2. If the cell concentration exceeds the required level, do not centrifuge, but adjust the concentration to between 2 x 106 and 2 x 107 cysts/mL with fresh medium. If the concentration is too low, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration.
  3. While cells are centrifuging, prepare a 15% (v/v) solution of sterile DMSO as follows: Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath. Allow the DMSO to solidify. Add the required volume of refrigerated medium. Dissolve the DMSO by inverting the tube several times.
    *NOTE: 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 between 106 and 107 cells/mL and 7.5% (v/v) DMSO. 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 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 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 sufficiently 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 712 in a T-25 tissue culture flask or plastic 16 x 125 mm screw-capped test tube. Incubate at 25°C.
Name of Depositor RJ Neff
Year of Origin 1957
References

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Cross References

Nucleotide (GenBank) : K00471 A.castellanii (amoeba) 5.8S ribosomal RNA.

Nucleotide (GenBank) : U12386 Acanthamoeba castellanii mitochondrion, complete genome.

Nucleotide (GenBank) : U03732 Acanthamoeba castellanii Neff mitochondrion rRNA large subunit gene.

Nucleotide (GenBank) : M97651 Acanthamoeba castellanii mitochondrial Met-tRNA, Ala-tRNA, Pro-tRNA, and Asp-tRNA genes.

Nucleotide (GenBank) : L20506 Acanthamoeba castellanii transfer RNA-Gln, complete tRNA; transfer RNA-Lys, complete tRNA; transfer RNA-Glu, complete tRNA; transfer RNA-Ile, complete tRNA; transfer RNA-Leu, complete tRNA.