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
Clone of strain WB (ATCC 30957), 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
Genome Sequenced Strain
Yes
Comments
Excellent cyst-formation under axenic conditions
Variable surface antigen
Differential mRNA processing
Genome sequencing strain
Medium
ATCC® Medium 2695: Keister's Modified TYI-S-33
ATCC® Medium 2155: LYI Giardia Medium (filtered)
Growth Conditions
Temperature: 35°C Culture System: Axenic
Cryopreservation
Harvest and Preservation
Harvest cells from a culture that is at or near peak density. To detach cells from the wall of the culture tubes place on ice for 10 minutes. Invert tubes several times until the majority of the cells are in suspension. Centrifuge tubes at 800 x g for 5 minutes.
Adjust the concentration of cells to 2 x 107/mL in fresh medium.
Before centrifuging prepare a 24% (v/v) solution of sterile DMSO in fresh medium containing 8% (w/v) sucrose. The solution is prepared as follows:
Add 1.05 g sucrose to 10 mL of fresh medium and filter sterilize through a 0.2 µm filter;
Add 2.4 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 7.6 mL of ice cold medium prepared in step 3a. The final concentration will be 24% (v/v) DMSO and 8% (w/v) sucrose;
Invert several times to dissolve the DMSO;
Allow to warm to room temperature.
Mix the cell preparation and the cryoprotective agent, prepared in step 3, in equal portions. Thus, the final concentration will equal 12% (v/v) DMSO + 4% sucrose (w/v) and 107 cells/mL. The time from the mixing of the cell preparation and DMSO stock solution to the start of the freezing process should be no less than 15 min and no longer than 30 min.
Dispense in 0.5 mL aliquots into 1.0 - 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.
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 13 mL ATCC Medium 2695.
Incubate the culture on a 15º horizontal slant at 35°C.
Name of Depositor
FD Gillin
Special Collection
NCRR Contract
Year of Origin
1983
References
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Aley SB, Gillin FD. Giardia lamblia: post-translational processing and status of exposed cysteine residues in TSA 417, a variable surface antigen. Exp. Parasitol. 77: 295-305, 1993. PubMed: 8224085
Aley SB, et al. Killing of Giardia lamblia by cryptdins and cationic neutrophil peptides. Infect. Immun. 62: 5397-5403, 1994. PubMed: 7960119
Das S, Gillin FD. Giardia lamblia: increased UDP-N-acetyl-D-glucosamine and N-acetyl-D-galactosamine transferase activities during encystation. Exp. Parasitol. 83: 19-29, 1996. PubMed: 8654548
Eckmann L, et al. Nitric oxide production by human intestinal epithelial cells and competition for arginine as potential determinants of host defense against the lumen-dwelling pathogen Giardia lamblia. J. Immunol. 164: 1478-1487, 2000. PubMed: 10640765
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McArthur AG, et al. The evolutionary origins of eukaryotic protein disulfide isomerase domains: new evidence from the Amitochondriate protist Giardia lamblia. Mol. Biol. Evol. 18: 1455-1463, 2001. PubMed: 11470836
Morrison HG, et al. Giardia lamblia expresses a proteobacterial-like DnaK homolog. Mol. Biol. Evol. 18: 530-541, 2001. PubMed: 11264404
Que X, et al. Developmentally regulated transcripts and evidence of differential mRNA processing in Giardia lamblia. Mol. Biochem. Parasitol. 81: 101-110, 1996. PubMed: 8892309
Reiner DS, et al. Giardia lamblia: absence of cyst antigens and reduced secretory vesicle formation and bile salt uptake in an encystation-deficient subline. Exp. Parasitol. 77: 461-472, 1993. PubMed: 7504633
Reiner DS, et al. A lipoprotein-cholesterol-albumin serum substitute stimulates Giardia lamblia encystation vesicle formation. J. Eukaryot. Microbiol. 42: 622-627, 1995. PubMed: 7581338
Sanchez LB, et al. Cloning and sequencing of an acetyl-CoA synthetase (ADP-forming) gene from the amitochondriate protist, Giardia lamblia. Gene 233: 225-231, 1999. PubMed: 10375639
Sun C-H, et al. A novel Myb-related protein involved in transcriptional activation of encystation genes in Giardia lamblia. Mol. Microbiol. 46: 971-984, 2002. PubMed: 12421304
