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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:
   1. Add 1.05 g sucrose to 10 mL of fresh medium and filter sterilize through a 0.2 µm filter;
   2. Add 2.4 mL of DMSO to an ice cold 20 x 150 mm screw-capped test tube;
   3. 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;
   4. Invert several times to dissolve the DMSO;
   5. 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 to the start of the freezing process 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 15º horizontal slant at 35°C.

Schupp DG, et al. Production of viable Giardia cysts in vitro: determination by fluorogenic dye staining, excystation, and animal infectivity in the mouse and Mongolian gerbil. Gastroenterology 95: 1-10, 1988. PubMed: 3286359

Abaza SM, et al. Isoenzyme profiles of four strains of Giardia lamblia and their infectivity to jirds. Am. J. Trop. Med. Hyg. 44: 63-68, 1991. PubMed: 1996742

Gillin FD, et al. Small-intestinal factors promote encystation of Giardia lamblia in vitro. Infect. Immun. 56: 705-707, 1988. PubMed: 3343054

Nash TE, et al. Restriction-endonuclease analysis of DNA from 15 Giardia isolates obtained from humans and animals. J. Infect. Dis. 152: 64-73, 1985. PubMed: 2409186

van Keulen H, et al. Cloning and restriction enzyme mapping of ribosomal DNA of Giardia duodenalis, Giardia ardeae and Giardia muris. Mol. Biochem. Parasitol. 46: 275-284, 1991. PubMed: 1922199

Kirkpatrick CE, Green GA. Susceptibility of domestic cats to infections with Giardia lamblia cysts and trophozoites from human sources. J. Clin. Microbiol. 21: 678-680, 1985. PubMed: 3998096

Werries E, et al. Purification and substrate specificity of two cysteine proteinases of Giardia lamblia. J. Protozool. 38: 378-383, 1991. PubMed: 1787424

Moss DM, et al. Isoenzyme comparison of axenic Giardia lamblia strains. J. Protozool. 39: 559-564, 1992. PubMed: 1522537

Reiner DS, et al. Sorting of cyst wall proteins to a regulated secretory pathway during differentiation of the primitive eukaryote, Giardia lamblia. Eur. J. Cell Biol. 53: 142-153, 1990. PubMed: 2076701

Gillin FD, et al. Giardia lamblia: the roles of bile, lactic acid, and pH in the completion of the life cycle in vitro. Exp. Parasitol. 69: 164-174, 1989. PubMed: 2753120

Campbell SR, et al. Giardia sp.: comparison of electrophoretic karyotypes. Exp. Parasitol. 71: 470-482, 1990. PubMed: 2226707

Faubert G, et al. Giardia lamblia: regulation of secretory vesicle formation and loss of ability to reattach during encystation in vitro. Exp. Parasitol. 72: 345-354, 1991. PubMed: 2026212

Isaac-Renton JL, et al. Comparison of an in vitro method and an in vivo method of Giardia excystation. Appl. Environ. Microbiol. 58: 1530-1533, 1992. PubMed: 1622221

Isaac-Renton JL, et al. Characterization of Giardia duodenalis isolates from a waterborne outbreak. J. Infect. Dis. 167: 431-440, 1993. PubMed: 8421176

Kirkpatrick CE, et al. Biochemical characterization of some raptor trypanosomes. II. Enzymes studies with a description of Trypanosoma bennetti new species. Can. J. Zool. 64: 195-203, 1986.

Soltys BJ, Gupta RS. Presence and cellular distribution of a 60-KDA protein related to mitochondrial HSP60 in Giardia lamblia. J. Parasitol. 80: 580-590, 1994. PubMed: 7914923

Rozario C, et al. Primary structure and phylogenetic relationships of glyceraldehyde-3-phosphate dehydrogenase genes of free-living and parasitic diplomonad flagellates. J. Eukaryot. Microbiol. 43: 330-340, 1996. PubMed: 8768438

Wallis PM, et al. Prevalence of Giardia cysts and Cryptosporidium oocysts and characterization of Giardia spp. isolated from drinking water in Canada. Appl. Environ. Microbiol. 62: 2789-2797, 1996. PubMed: 8702271

Gillin FD, et al. Isolation and expression of the gene for a major surface protein of Giardia lamblia. Proc. Natl. Acad. Sci. USA 87: 4463-4467, 1990. PubMed: 2352929

Nash TE, et al. Excretory-secretory products of Giardia lamblia. J. Immunol. 131: 2004-2010, 1983. PubMed: 6619547

Ong C, et al. Studies of Giardia spp. and Cryptosporidium spp. in two adjacent watersheds. Appl. Environ. Microbiol. 62: 2798-2805, 1996. PubMed: 8702272

Baum KF, et al. Purine nucleoside and nucleobase cell membrane transport in Giardia lamblia. J. Eukaryot. Microbiol. 40: 643-649, 1993. PubMed: 8401476

Soltys BJ, Gupta RS. Immunoelctron microscopy of Giardia lamblia cytoskeleton using antibody to acetylated gamma-tubulin. J. Eukaryot. Microbiol. 41: 625-632, 1994. PubMed: 7866386

Aley SB, et al. Killing of Giardia lamblia by cryptdins and cationic neutrophil peptides. Infect. Immun. 62: 5397-5403, 1994. PubMed: 7960119

Johns T, et al. Anti-giardial activity of gastroinstestihnal remedies of the Luo of East Africa. J. Ethnopharmacol. 46: 17-23, 1995. PubMed: 7475119

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

McCabe RE, et al. In vitro model of attachment of Giardia intestinalis trophozoites to EIC-6 cells, an intestinal cell line. Antimicrob. Agents Chemother. 35: 29-35, 1991. PubMed: 1901700

Bell CA, et al. Structure-activity relationships of pentamidine analogs against Giardia lamblia and correlation of antigardial activity with DNA-building affinity. Antimicrob. Agents Chemother. 35: 1099-1107, 1991. PubMed: 1929249

Gillin FD, et al. Human milk kills parasitic intestinal protozoa. Science 221: 1290-1292, 1983. PubMed: 6310751

Wieder SC, et al. Mass cultivation of Giardia lamblia in a serum-free medium. J. Parasitol. 69: 1181-1182, 1983. PubMed: 6674471

Campbell JD, Faubert GM. Comparative studies on Giardia lamblia encystation in vitro and in vivo. J. Parasitol. 80: 36-44, 1994. PubMed: 8308656

Gillin FD. Giardia lamblia: the role of conjugated and unconjugated bile salts in killing by human milk. Exp. Parasitol. 63: 74-83, 1987. PubMed: 3803534

Reiner DS, et al. Identification and localization of cyst-specific antigens of Giardia lamblia. Infect. Immun. 57: 963-968, 1989. PubMed: 2917795

Baum KF, et al. Purine deoxynucleoside salvage in Giardia lamblia. J. Biol. Chem. 264: 21087-21090, 1989. PubMed: 2480350

Gillin FD, et al. Biliary lipids support serum-free growth of Giardia lamblia. Infect. Immun. 53: 641-645, 1986. PubMed: 3744557

Reiner DS, Gillin FD. Human secretory and serum antibodies recognize environmentally induced antigens of Giardia lamblia. Infect. Immun. 60: 637-643, 1992. PubMed: 1730497

Hou G, et al. Structure of a frequently rearranged rRNA-encoding chromosome in Giardia lamblia. Nucleic Acids Res. 23: 3310-3317, 1995. PubMed: 7667108

Smith PD, et al. Chronic giardiasis: studies on drug sensitivity, toxin production, and host immune response. Gastroenterology 83: 797-803, 1982. PubMed: 7106510

Moss DM, et al. Antigenic variation of Giardia lamblia in the feces of Mongolian gerbils. J. Clin. Microbiol. 28: 254-257, 1990. PubMed: 2312672

Torian BE, et al. Specific and common antigens of Trichomonas vaginalis detected by monoclonal antibodies. Infect. Immun. 43: 270-275, 1984. PubMed: 6360900

Dickerson JW, et al. Infectivity of cryopreserved Giardia cysts for Mongolian gerbils (Meriones unguiculatus). J. Parasitol. 77: 688-691, 1991. PubMed: 1919914

Gillin FD, et al. Inhibition of growth of Giardia lamblia by difluoromethylornithine, a specific inhibitor of polyamine biosynthesis. J. Protozool. 31: 161-163, 1984. PubMed: 6330350

Moss DM, et al. Purification and characterization of Giardia lamblia antigens in the feces of Mongolian gerbils. J. Clin. Microbiol. 29: 21-26, 1991. PubMed: 1704383

Udezulu IA, et al. Isolation of two Giardia lamblia (WB strain) clones with distinct surface protein and antigenic profiles and differing infectivity and virulence. Infect. Immun. 60: 2274-2280, 1992. PubMed: 1587594

Erlandsen SL, et al. Axenic culture and characterization of Giardia ardeae from the great blue heron (Ardea herodias). J. Parasitol. 76: 717-724, 1990. PubMed: 2213415

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

Hill DR, Pohl R. Ingestion of Giardia lamblia trophozoites by murine Peyer's patch macrophages. Infect. Immun. 58: 3202-3207, 1990. PubMed: 2401561

Meng TC, et al. Inhibition of Giardia lamblia excystation by antibodies against cyst walls and by wheat germ agglutinin. Infect. Immun. 64: 2151-2157, 1996. PubMed: 8675320

Feely DE, Dyer JK. Localization of acid phosphatase activity in Giardia lamblia and Giardia muris trophozoites. J. Protozool. 34: 80-83, 1987.

Keeling PJ, et al. Evolutionary relationship between translation initiation factor eIF-2gamma and selenocysteine-specific elongation factor SELB: change of function in translation factors. J. Mol. Evol. 47: 649-655, 1998. PubMed: 9847405

Edgell DR, et al. Evidence of independent gene duplications during the evolution of archael and eukaryotic family B DNA polymerases. Mol. Biol. Evol. 15: 1207-1217, 1998. PubMed: 9729885

Sousa MC, Poiares-Da-Silva J. A new method for assessing metronidazole susceptibility of Giardia lamblia trophozoites. Antimicrob. Agents Chemother. 43: 2939-2942, 1999. PubMed: 10582886

Sousa MC, et al. Adherence of Giardia lamblia trophozoites to Int-407 human intestinal cells. Clin. Diagn. Lab. Immunol. 8: 258-265, 2001. PubMed: 11238205

Perez PF, et al. Inhibition of Giardia intestinalis by extracellular factors from Lactobacilli: an in vitro study. Appl. Environ. Microbiol. 67: 5037-5042, 2001. PubMed: 11679323

Archibald JM, et al. Origin and evolution of eukaryotic chaperonins: phylogenetic evidence for ancient duplications in CCT genes. Mol. Biol. Evol. 17: 1456-1466, 2000. PubMed: 11018153

Arguello-Garcia A, et al. Sequential exposure and assembly of cyst wall filaments on the surface of encysting Giardia duodenalis. Parasitology 125: 209-219, 2002. PubMed: 12358418

Yang H-W, et al. Characterization of two glyceraldehyde 3-phosphate dehydrogenase genes in Giardia lamblia. Parasitol. Res. 88: 646-650, 2002. PubMed: 12107457

Sousa MC, Poiares-da-Silva J. The cytotoxic effects of ciprofloxacin in Giardia lamblia trophozoites. Toxicol. In Vitro 15: 297-301, 2001. PubMed: 11566552

Miller RL, et al. Identification of Giardia lamblia Isolates Susceptible and Resistant to Infection by the Double-Stranded RNA Virus. Exp. Parasitol. 66: 118-123, 1988. PubMed: 3366209

Nucleotide (GenBank) : U24238 Giardia lamblia (ENC1) mRNA, complete cds.

Nucleotide (GenBank) : U24239 Giardia lamblia (ENC6) mRNA, complete cds.

Nucleotide (GenBank) : U42428 Giardia lamblia caltractin (CAL) gene, complete cds.

Nucleotide (GenBank) : AF067402 DNA polymerase alpha gene sequence, partial coding sequence

Nucleotide (GenBank) : U19901 Giardia lamblia putative adenylate kinase gene, complete cds.

Nucleotide (GenBank) : U10907 Giardia lamblia WB putative surface antigen gene, partial cds.

Nucleotide (GenBank) : X60427 G.lamblia telomere DNA containing truncated rRNA large subunit gene.

Nucleotide (GenBank) : M63966 Giardia lamblia variant-specific surface protein 1267 (vsp1267) gene, complete cds.

Nucleotide (GenBank) : AF005078 translation initiation factor 2 gamma subunit, eIF-2 gamma gene, partial coding sequence

Nucleotide (GenBank) : U12337 Giardia lamblia WB inorganic pyrophosphate-linked phosphofructokinase (GLPFK) gene, complete cds.