Trichomonas vaginalis Donne (ATCC® 50147)

Organism: Trichomonas vaginalis Donne  /  Depositor: M Muller

Strain Designations NYH 272
Application
Sexually Transmitted Disease Research
Biosafety Level 2
Isolation
human, New York Hospital, New York City, 1977
Product Format frozen
Type Strain no
Comments
Metronidazole sensitive.
Growth Conditions
Temperature: 35.0°C
Duration: axenic; anaerobic
Protocol: at pH 6.0
Subcultivation
Protocol: at pH 6.0
Cryopreservation

1.  Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min. The cells grown in a medium containing agar are concentrated by centrifugation, a solid pellet does not form. The soft pellet is resuspended to desired cell concentration with agar-free supernatant.

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

3.  While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium.

a) Add 1.0 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 9.0 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 cells/ml and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should 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. Vials should not be stored above -55°C.

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

9.   Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate a 16 x 125 mm screw-capped test tube containing 13 mls of ATCC medium 1978 adjusted to pH 6.0.

10.          Incubate the culture on a 15° horizontal slant at 35°C.

 

Name of Depositor M Muller
Year of Origin 1977
References

Muller M, Gorrell TE. Metabolism and metronidazole uptake in Trichomonas vaginalis isolates with different metronidazole susceptibilities. Antimicrob. Agents Chemother. 24: 667-673, 1983. PubMed: 6607028

Alderete JF. Antigen analysis of several pathogenic strains of Trichomonas vaginalis. Infect. Immun. 39: 1041-1047, 1983. PubMed: 6601618

Muller M. Mode of action of metronidazole on anaerobic bacteria and protozoa. Surgery 93: 165-171, 1983. PubMed: 6849201

Muller M, et al. Three metronidazole-resistant strains of Trichomonas vaginalis from the United States. Am. J. Obstet. Gynecol. 138: 808-812, 1980. PubMed: 7004190