Tetrahymena thermophila Nanney and McCoy (ATCC® 30007)

Strain Designations: WH-6 (WHI) [ATCC 16539]  /  Depositor: AM Elliott  /  Biosafety Level: 1

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Deposited As Tetrahymena pyriformis (Ehrenberg) Lwoff
Strain Designations WH-6 (WHI) [ATCC 16539]
Application
characterization
Identification of Tetrahymena species using PCR/RFLP analysis of rDNA
Biosafety Level 1
Isolation
freshwater pond, Woods Hole, MA, 1952
Product Format test tube
Type Strain yes
Comments
I
characterization
Identification of Tetrahymena species using PCR/RFLP analysis of rDNA
Cell volume distributions during the growth
A biometric study
Inhibition of conjugation and cell division
Medium Medium 357: Tetrahymena medium
Growth Conditions
Temperature: 25.0°C
Duration: axenic
Cryopreservation
1.  Transfer Tetrahymena from usual growth medium to ATCC Medium 1034 and allow to grow to near peak density.

2.   Harvest cells from a culture by centrifugation at 300 x g for 2 min.          

3.   Adjust concentration of cells to 2 x 106/ml in fresh

      medium.

4.   While cells are centrifuging, prepare a 22% (v/v) sterile

solution of sterile DMSO in fresh medium.

a) Add 2.2 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 7.8 ml of ice cold medium;

c) Invert several times to dissolve the DMSO;

d) Allow to warm to room temperature.

5.   Add a volume of the DMSO solution equal to the cell

      suspension volume but add in 3 equal aliquots at 2 min

      intervals. Thus, the final concentration of the preparation

      will equal 11% (v/v) DMSO and 106 cells /ml.

6.   Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic

      screw-capped cryules (special plastic vials for       cryopreservation).

7.   Place the ampules in a controlled rate freezing unit. The

cooling cycle should be initiated no less than 15 min and no longer than 60 min after the addition of the DMSO to the cell preparation. 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  -50°C ampules are plunged into liquid nitrogen.

8.   Store in the vapor or liquid phase of a nitrogen

      refrigerator.

9.   To establish a culture from the frozen state aseptically add 0.5 ml of sterile modified PYNFH medium (ATCC Medium 1034) containing 8% (w/v) sucrose to the ampule.  Immediately, place in a 35°C water bath, until thawed. Immerse the ampule just sufficient to cover the frozen material. Do not agitate the ampule.

10.          Immediately after thawing, aseptically remove the contents of the ampule and gently add the material to the edge of a 20 x 100 mm Petri plate containing ATCC Medium 919 (non-nutrient agar) and position on a 15 degree slant.  The cell suspension will pool at the edge of the plate.

11.          Continue to double the volume of the cell suspension at 10

minute intervals by adding ATCC medium 1034) containing 4% sucrose (w/v).  When the volume reaches 16.0 ml place the plate in horizontal position and incubate at 25°C. 

12.          On the following day, gently remove the cell suspension for the plate and transfer to a T-25 tissue culture flask.  Note the volume of the suspension and add a volume of fresh medium containing 4% sucrose equal to the volume of  the cell suspension.  Incubate the culture at 25°C.

13.          After culture has been established subculture into fresh

      normal medium without sucrose. 

Name of Depositor AM Elliott
Chain of Custody
ATCC <<--AM Elliott<<--A. Elliott/D.F. Gruchy
Year of Origin 1952
References

. . Annu. Rev. Microbiol. 13: 79-96, 1959.

Biol. Bull. 105: 269-284, 1953.

Jerome CA, Lynn DH. Identifying and distinguishing sibling species in the Tetrahymena pyriformis complex (Ciliophora, Oligohymenophora) using PCR/RFLP analysis of nuclear ribosomal DNA. J. Eukaryot. Microbiol. 43: 492-497, 1996. PubMed: 8976607

Hallewell RA, Mullenbach GT. DNA encoding human cytoplasmic Cu/Zn superoxide dismutase. US Patent 5,629,189 dated May 13 1997

Gates MA, Berger J. Morphologic stability in Tetrahymena pyriformis. Trans. Am. Microsc. Soc. 95: 11-22, 1976. PubMed: 817430

Gates MA. Trajectories of cell volume distributions during the growth cycle of Tetrahymena. J. Gen. Microbiol. 129: 895-900, 1983.

Holz GG, et al. The arrest of mitosis and stomatogensis during temperature-induction of synchronous division in Tetrahymena pyriformis, mating type 1, variety 1. Exp. Cell Res. 13: 618-621, 1957. PubMed: 13490386

Frisch A, et al. Inhibition of conjugation and cell division in Tetrahymena pyriformis by tunicamycin: A possible requirement of glycoprotein synthesis for induction of conjugation. Biochem. Biophys. Res. Commun. 72: 138-145, 1976. PubMed: 825115

Bleyman LK, Simon EM. Genetic control of maturity in Tetrahymena pyriformis. Genet. Res. 10: 319-321, 1967. PubMed: 5587947

Gates MA, Berger J. A biometric study of three strains of Tetrahymena pyriformis (Ciliatea: Hymenostomatida). Can. J. Zool. 52: 1167-1183, 1974. PubMed: 4214189

Nanney DL, McCoy JW. Characterization of the species of the Tetrahymena pyriformis complex. Trans. Am. Microsc. Soc. 95: 664-682, 1975.

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