Giardia intestinalis (Lambl) Alexeieff (ATCC® 50582)

Organism: Giardia intestinalis (Lambl) Alexeieff  /  Depositor: TE Nash

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Deposited As Giardia lamblia (Lambl) Stiles
Strain Designations WB clone 1267
Application
Enteric Research
Food and waterborne pathogen research
Biosafety Level 2
Product Format frozen
Type Strain no
Comments
Serum Cohn Fraction IV-1 supports growth
Triosephosphate isomerase gene
Metabolism of cysteine
Isoprenylation of proteins
Induction of Golgi
Lipid requirements
Induction of differentiation by cholesterol starvation
Expression of cyst wall protein gene
Medium ATCC® Medium 2155: LYI Giardia Medium (filtered)
Growth Conditions
Temperature: 35.0°C
Duration: axenic; anaerobic
Cryopreservation

1.   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. 

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

3.  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:

a) Add 10.5 g sucrose to 10 ml of fresh medium and filter sterilize through a 0.2 mm filter;

b) Add 2.4 ml of DMSO to an ice cold 20 x 150 mm screw-capped test tube;

c) 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;

d) Invert several times to dissolve the DMSO;

e) Allow to warm to room temperature.

4.   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 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.

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 ml ATCC Medium 2695.

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

Name of Depositor TE Nash
Special Collection NCRR Contract
References

Lujan HD, et al. Serum Cohn fraction IV-1 supports the growth of Giardia lamblia in vitro. Infect. Immun. 62: 4664-4666, 1994. PubMed: 7927737

Mowatt MR, et al. Complementation of an Escherchia coli glycolysis mutant by Giardia lamblia triosephosphate isomerase. Exp. Parasitol. 78: 85-92, 1994. PubMed: 8299763

Lujan HD, Nash TE. The uptake and metabolism of cysteine by Giardia lamblia trophozoites. J. Eukaryot. Microbiol. 41: 169-175, 1994. PubMed: 8167619

Lujan HD, et al. Isoprenylation of proteins in the protozoan Giardia lamblia. Mol. Biochem. Parasitol. 72: 121-127, 1995. PubMed: 8538683

Lujan HD, et al. Developmental induction of Golgi structure and function in the primitive eukaryote Giardia lamblia. J. Biol. Chem. 270: 4612-4618, 1995. PubMed: 7876232

Lujan HD, et al. Lipid requirements and lipid uptake by Giardia lamblia trophozoites in culture. J. Eukaryot. Microbiol. 43: 237-242, 1996. PubMed: 8640194

Lujan HD, et al. Cholesterol starvation induces differentiation of the intestinal parasite Giardia lamblia. Proc. Natl. Acad. Sci. USA 93: 7628-7633, 1996. PubMed: 8755526

Mowatt MR, et al. Developmentally regulated expression of a Giardia lamblia cyst wall protein gene. Mol. Microbiol. 15: 955-963, 1995. PubMed: 7596296

Lujan HD, et al. Increased expression of the molecular chaperone BiP/GRP78 during the differentiation of a primitive eukaryote. Biol. Cell. 86: 11-18, 1996. PubMed: 8688827

Nash T, Rice WG. Efficacies of zinc-finger-active drugs against Giardia lamblia. Antimicrob. Agents Chemother. 42: 1488-1492, 1998. PubMed: 9624499

Elmendorf HG, et al. The abundance of sterile transcripts in Giardia lamblia. Nucleic Acids Res. 29: 4674-4683, 2001. PubMed: 11713317

Touz MC, et al. Identification and characterization of a novel secretory granule calcium-binding protein from the early branching eukaryote Giardia lamblia. J. Biol. Chem. 277: 50557-50563, 2002. PubMed: 12354777

Touz MC, et al. The activity of a developmentally regulated cysteine proteinase is required for cyst wall formation in the primitive eukaryote Giardia lamblia. J. Biol. Chem. 277: 8474-8481, 2002. PubMed: 11773053

Lujan HD, et al. Identification of a novel Giardia lamblia cyst wall protein with leucine-rich repeats. Implications for secretory granule formation and protein assembly into the cyst wall. J. Biol. Chem. 270: 29307-29313, 1995. PubMed: 7493963

Murtagh JJ Jr., et al. Guanine nucleotide-binding proteins in the intestinal parasite Giardia lamblia. Isolation of a gene encoding an approximately 20-kDa ADP-ribosylation factor. J. Biol. Chem. 267: 9654-9662, 1992. PubMed: 1577802

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.
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