Salpingoeca napiformis Kent (ATCC® 50153)

Organism: Salpingoeca napiformis Kent  /  Depositor: TA Nerad

Permits and Restrictions

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Deposited As Salpingoeca sp.
Strain Designations ATCC:0585:1
Biosafety Level 1
Isolation
Biomphalaria glabrata aquarium culture, Rockville, MD, 1985
Product Format frozen
Type Strain no
Medium ATCC® Medium 802: Sonneborn's Paramecium medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with Enterobacter aerogenes ATCC 13048 and mixed bacteria
Cryopreservation
1.   Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.

2.  Adjust the concentration of cells to 2 x 106 - 107/ml in fresh medium.

3.  While cells are centrifuging prepare a 20% (v/v) solution of sterile DMSO in fresh medium.

a) Add 2.0 ml of DMSO to an ice cold 20 x 150 mm screw-capped test tube;

b) Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 8.0 ml of ice cold medium;

c) Invert several times to dissolve the DMSO;

d) Allow to warm to room temperature.

4.  Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be 106 - 107 and 10% (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 30 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 should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. If frozen material is to be stored at temperatures between -130°C and -70°C the shelf life should be empirically tested, i.e., remove stored material at intervals to determine die-off rate.

8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the ampule to a level just above the surface of the frozen material. Do not agitate the ampule.

9. Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate a T-25 tissue culture flask containing 10 ml of bacterized ATCC medium 802.

10.          Incubate with the cap screwed on tightly at 25°C.

Disclaimers

This product is intended for laboratory research purposes only. It is not intended for use in humans.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate.

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Please see the enclosed Material Transfer Agreement (MTA) for further details regarding the use of this product. The
MTA is also available on our Web site at www.atcc.org.

© ATCC 2003. All rights reserved.

ATCC® is a registered trademark of the American Type Culture Collection.

1/03 TAN

Name of Depositor TA Nerad
Year of Origin 1985
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