Naegleria gruberi Schardinger (ATCC® 30133)

Strain Designations: EG  /  Depositor: W Balamuth  /  Biosafety Level: 1

Permits and Restrictions

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Strain Designations EG
Biosafety Level 1
Isolation
soil in a eucalyptus grove, Berkeley, CA, 1959
Product Format frozen
Type Strain no
Comments
Electron microscope study
High-resolution (PGGE) of isoenzymes
Medium Medium 997: Fresh water ameba medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with Escherichia coli ATCC 11775
Cryopreservation
Cryoprotective Solution

DMSO                                                                                    1.5 ml

Page's Balanced Salt 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 1323 (Page's Balanced Salt 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.

10.   Follow the protocol for maintenance of culture.

Name of Depositor W Balamuth
Chain of Custody
ATCC <<--W Balamuth<<--F.L. Schuster
Year of Origin 1959
References

Fulton C. Early events of cell differentiation in Naegleria gruberi. Synergistic control by electrolytes and a factor from yeast extract. Dev. Biol. 28: 603-619, 1972. PubMed: 5049526

Schuster F. An electron microscope study of the amoebo-flagellate, Naegleria gruberi (Schardinger). I. The amoeboid and flagellate stages. J. Protozool. 10: 287-313, 1963.

Moss DM, et al. High-resolution polyacrylamide gradient gel electrophoresis (PGGE) of isoenzymes from five Naegleria species. J. Protozool. 35: 26-31, 1988. PubMed: 2966859

De Jonckheere JF. A century of research on the amoeboflagellate genus Naegleria. Acta Protozool. 41: 309-342, 2002.

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