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Toxoplasma gondii (Nicolle and Manceaux) Nicolle and Manceaux 菌株拉丁名

(ATCC? 50951?) 菌株編號 

Deposited As Toxoplasma gondii (Nicolle and Manceaux) Nicolle and Manceaux 

Strain Designations 菌株別名 C56 

Application 用途 Food and waterborne pathogen research Opportunistic pathogen research 

Biosafety Level 生物安全等級 2 

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 分離源 Clinical specimen - animal, chicken with chronic infection, USA 

Product Format 提供形式 frozen 

Storage Conditions 保藏方法 

Frozen Cultures 冷凍物 : -70°C for 1 week; liquid N2 vapor for long term storage  

Freeze-dried  Cultures 凍干物 : 2-8°C 

Live Cultures 活菌 : See Protocols section for handling information 

Type Strain 模式菌株 no 

Genotype Haplogroup 3

Growth Conditions 生長條件

Temperature 培養(yǎng)溫度 : 35°C to 37°C 

Cell Line: ATCC? CRL-1634? (human foreskin fibroblasts) 

Cryopreservation Harvest and Preservation 

1.To harvest the Toxoplasma culture, detach any remaining tissue culture cells (infected and uninfected) by scraping the surface of the flask with a cell scraper. 

2.Transfer the cell suspension (including parasites) to 15 mL plastic centrifuge tubes. Centrifuge at 1300 x g for 10 min. 

3.Remove all but 0.5 mL of the supernatant from each tube, resuspend the cell pellets, and pool them to a single tube. 

4.Pass the resulting cell suspension through a syringe equipped with a 27 gauge 1/2 in needle to break up any remaining cells. Adjust the parasite concentration to 2.0 - 4.0 x 107 cells/mL with fresh medium or PBS. 

NOTE: If the concentration of parasites is too low, centrifuge at 1300 x g for 10 min and resuspend in the volume of fresh medium or PBS required to yield the desired concentration. 

5.Prepare a cryoprotective solution containing 15% (v/v) DMSO and 50% (v/v) HIFBS in fresh medium or PBS. 

6.Mix the cell preparation and cryoprotective solution in equal portions. The final concentration will be 1.0 - 2.0 x 107 cells/mL, 7.5% DMSO, and 25% HIFBS. The time from the mixing of the cell preparation and cryoprotective solution to the start of the freezing process should be no less than 15 min. and no more than 30 min. 

NOTE: To prevent culture contamination, penicillin-streptomycin solution (ATCC? 30-2300) may be added to a final concentration of 50 to 100 I.U./mL penicillin and 50 to 100 μg/mL streptomycin. 

7.Dispense in 0.5 mL aliquots to 1.0-2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 

8.Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through heat of fusion. 

At -40°C plunge ampules 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.) 

9.Store frozen ampules in either the vapor or liquid phase of a nitrogen refrigerator. 

10.To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after thawed. 

11.Immediately after thawing, aseptically transfer contents to a T-25 tissue culture flask containing a fresh monolayer of ATCC? CRL-1634? cells and 10 mL ATCC? 30-2002 with 3% (v/v) HIFBS. 

12.Outgas the flask for 10 seconds with a 95% air, 5% CO2 gas mixture. 

13.Incubate in a 35-37°C CO2 incubator with the cap screwed on tightly. Mycoplasma NoName of Depositor LD Sibley, J Remington Special Collection NCRR Contract 

References 參考文獻 Howe DK, Sibley LD. Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease. J. Infect. Dis. 172: 1561-1566, 1995. PubMed: 7594717 

Sibley LD, Boothroyd JC. Virulent strains of Toxoplasma gondii comprise a single clonal lineage. Nature 359: 82-85, 1992. PubMed: 1355855 

clonal lineage [50660] [50661] Asai T, et al. Biochemical and molecular characterization of nucleoside triphosphate hydrolase isozymes from the parasitic protozoan Toxoplasma gondii. J. Biol. Chem. 270: 11391-11397, 1995. PubMed: 7744775 L D Sibley, personal communication