Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Isolation
Human, Massachusetts, 1983
Product Format
frozen
Storage Conditions
Frozen Cultures: -70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures: 2-8°C
Live Cultures: See Protocols section for handling information
Type Strain
no
Comments
In vitro susceptibility of metronidazole
Medium
ATCC® Medium 2154: LYI Entamoeba medium
ATCC® Medium 361: Modified TYM basal medium (ATCC medium 358) with pH adjusted to 6.0 and 0.2-0.5 ml of heat-inactivated horse serum added per tube before use
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.
Adjust the concentration of cells
to 2 x 106 to 2 x 107/mL in fresh medium.
While cells
are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium.
Add 1.0 mL of DMSO to an ice cold 20 x 150 mm screw-capped test
tube.
Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 9.0 mL of
ice cold medium.
Invert several times to dissolve the DMSO.
Allow to warm to room
temperature.
Mix the cell preparation and the DMSO in equal portions. Thus, the
final concentration will be 106 to 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.
Dispense in 0.5 mL aliquots
into 1.0 mL to 2.0 mL sterile plastic screw-capped cryules (special plastic vials for
cryopreservation).
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.)
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.
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.
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 either 9 mL of ATCC medium 361 (completed
with serum) or 13 mL ATCC Medium 2154 adjusted to pH 6.0.
Incubate the culture at 35°C
with the cap screwed on tightly (tube should be vertical for medium 361 or on a 15° horizontal
slant for medium 2154).
Name of Depositor
M Muller
Chain of Custody
ATCC <-- M Muller <-- Hurd
Year of Origin
1983
References
Muller M. Mode of action of metronidazole on anaerobic bacteria and protozoa. Surgery 93: 165-171, 1983. PubMed: 6849201
Muller M, et al. In vitro susceptibility of Trichomonas vaginalis to metronidazole and treatment outcome in vaginal trichomoniasis. Sex. Transm. Dis. 15: 17-24, 1988. PubMed: 3258675
Cornelius DC, et al. Short report: genetic relatedness of Trichomonas vaginalis reference and clinical isolates. Am. J. Trop. Med. Hyg. 83: 1283-1286, 2010.
