Acanthamoeba castellanii (Douglas) Page 拉丁名
(ATCC® 50370™) 統(tǒng)一編號
Strain Designations 別名 Ma
Application 用途 Biochemical and molecular characterization This strain carries a potential mycobacterial endosymbiont based on culture in selective media and molecular analysis.
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 Human eye infection, New York, NY, 1978
Product Format 提供形式 frozen
Storage Conditions 保藏條件
Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Comments 注釋 Subgenus systematics based upon SSU rDNA sequence data Biochemical and molecular characterization in vitro effectiveness of povidone-iodine
This strain carries a potential mycobacterial endosymbiont based on culture in selective media and molecular analysis. Isolation Human eye infection, New York, NY, 1978
Medium 培養(yǎng)基 ATCC® Medium 712: PYG w/ Additives
Growth Conditions 生長條件
Temperature 培養(yǎng)溫度: 25°C
Culture System: Axenic
Cryopreservation Harvesting and Preservation
1,To achieve the best results set up cultures with several different inocula (e.g. 0.25 mL, 0.5 mL, 1.0 mL). Harvest cultures and pool when the culture that received the lowest inoculum is at or near peak density.
2， If the cell concentration exceeds the required level do not centrifuge, but adjust the concentration to between 2 x 106 and 2 x 107cysts/mL with fresh medium. If the concentration is too low, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration.
3,While cells are centrifuging prepare a 15% (v) solution of sterile DMSO as follows: Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath. Allow the DMSO to solidify. Add the required volume of refrigerated medium. Dissolve the DMSO by inverting the tube several times. Note: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
4，Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/mL and 7.5% (v
) DMSO. 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 mL to 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 are stored in either the vapor or liquid phase of a nitrogen freezer.,
8，To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min).
9， Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial.
Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5 mL of fresh ATCC medium 712 in a T-25 tissue culture flask or plastic 16 x 125 mm screw-capped test tube. Incubate at 25°C.
Name of Depositor寄存者 TJ Byers, RJ Gast
Chain of Custody 來源國家 ATCC <-- TJ Byers, RJ Gast <-- E Willaert <-- P Ma
Year of Origin收藏年份 1978
References 參考文獻 Chung DI, et al. Biochemical and molecular characterization of a strain KA/S2 of Acanthamoeba castellanii isolated from Korean soil. Korean J. Parasitol. 34: 79-85, 1996. PubMed: 8820744
Gast RJ, et al. Subgenus systematics of Acanthamoeba: Four nuclear 18S rDNA sequence types. J. Eukaryot. Microbiol. 43: 498-504, 1996. PubMed: 8976608
Gatti S, et al. In vitro effectiveness of povidone-iodine on Acanthamoeba isolates from human cornea. Antimicrob. Agents Chemother. 42: 2232-2234, 1998. PubMed: 9736540
Ledee DR, et al. Advantages of using mitochondrial 16S rDNA sequences to classify clinical isolates of Acanthamoeba. Invest. Ophthalmol. Vis. Sci. 44: 1142-1149, 2003. PubMed: 12601042
Kong HH, et al. Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small-subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. J. Clin. Microbiol. 40: 1199-1206, 2002. PubMed: 11923331
Robert E. Molestina, personal communication
Stothard DR, et al. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J. Eukaryot. Microbiol. 45: 45-54, 1998. PubMed: 9495032
Stothard DR, et al. Fluorescent oligonucleotide probes for clinical and environmental detection of Acanthamoeba and the T4 18S rRNA gene sequence type. J. Clin. Microbiol. 37: 2687-2693, 1999. PubMed: 10405422
Tachibana H, et al. Differentiation of Entamoeba histolytica from E. dispar facilitated by monoclonal antibodies against a 150-kDa surface antigen. Parasitol. Res. 83: 435-439, 1997. PubMed: 9197389
Cross References Nucleotide (GenBank) : U07414 nuclear SSU rRNA gene
