供货周期: | -- |
品牌: | 百欧博伟生物 |
规格: | 冻干粉 |
货号: | bio-109081 |
CAS号: | ATCC 30458 |
平台编号:Bio-109081
Entamoeba histolytica Schaudinn 拉丁名
ATCC 30458 编号
Permits and Restrictions
View Permits
Strain Designations 200:NIH
Application 用途 Enteric Research
Food and waterborne pathogen research
Biosafety Level 安全等级 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 Colonic biopsy from adult human male with amebic dysentery, passaged through dog and rabbit before establishment in culture, 1949
Product Format 提供形式 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
Comments
Axenic cultivation
Metronidazole metabolism
Zymodeme II and Mycoplasma-negative by PCR
Entamoeba phylogeny
Inhibition of growth by difluoromethyl-ornithine
Requirement for cysteine and ascorbic acid for attachment
Use of plastic ampoules for freeze preservation
Medium 培养基 ATCC Medium 2154: LYI Entamoeba medium
Growth Conditions 培养条件
Temperature 培养温度 : 35°C
Atmosphere 需氧情况 : Anaerobic
Culture System: Axenic
Cryopreservation Reagents
CPMB-5 Cryoprotective Solution
DMSO: 1.0 mL
2.5 M Sucrose: 0.8 mL
L-Cysteine/Ascorbic Acid Solution: 0.2 mL
CPMB-2 Basal Solution: 6.0 mL
HIBS: 2.0 mL
CPMB-2 Basal Solution
Yeast Extract: 60.0 g
K2HPO4: 1.0 g
KH2PO4: 0.6 g
NaCl: 2.0 g
Distilled water: 1.0 L
Autoclave for 15 minutes.
L-Cysteine/Ascorbic Acid Solution
L-Cysteine-HCL: 1.0 g
Acorbic Acid: 0.1 g
Distilled water: 10.0 mL
Add 9.0 mL of distilled water to a 20 mL beaker and dissolve the first two components. While stirring, adjust the pH to 7.2 with 10N NaOH (approximately 0.7 mL). Adjust final volume to 10 mL with distilled water and filter sterilize. Solution should be used soon after preparation. Discard any unused solution.
Harvest and Preservation
Harvest cells from several cultures that are in the late logarithmic to early stationary phase of growth. Place culture vessels on ice for 10 min.
Invert tubes 20 times and centrifuge at 200 x g for 5 min.
While cells are centrifuging, prepare the cryoprotective solution.
Place 1.0 mL DMSO in a 16 x 125 mm screw-capped test tube and ice until solidified. Add 0.8 mL of the 2.5 M Sucrose solution, remove from ice and invert until the DMSO is liquefied. Return to ice bath.
Add 0.2 mL of the L-Cysteine/Ascorbic Acid Solution to the DMSO solution and mix.
Add 6.0 mL of the CPMB-2 Basal solution and mix.
Add 2.0 mL HIBS and mix.
Resuspend the cell pellets and pool to a final volume of approximately 10 mL with the supernatant. Make a determination of the cell density and adjust the concentration of the cells between 5 x 105/mL and 1 x 106/mL using fresh medium. If the cell concentration is below 5 x 105/mL, centrifuge the cell suspension and resuspend the pellet in a volume that will yield the desired concentration.
After the cell concentration is adjusted, centrifuge as in step 2.
Remove as much supernatant as possible and determine the volume removed.
Resuspend the cell pellet with a volume of the cryoprotective solution equal to the volume of the supernatant removed. Invert the tube several times to obtain a uniform cell density.
Dispense 0.5 mL aliquots into 1.0 mL to 2.0 mL plastic sterile cryules (special plastic vials for cryopreservation).
Place the vials in a controlled rate freezing unit. Use the following cooling cycle:
From room temperature cool at -10°C/min to the heat of fusion.
From the heat of fusion to -40°C, cool at -1°C/min.
At -40°C plunge into liquid nitrogen. The cooling cycle should be initiated no less than 15 and no more than 30 minutes after the addition of DMSO to the cell preparation.
Store ampules in a liquid nitrogen refrigerator until needed.
To establish a culture from the frozen state, place an ampule in a 35°C water bath, until thawed (2 to 3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the ampule.
Transfer contents of thawed ampule to a 16 x 125 mm screw-capped borosilicate glass test tube containing 13 mL of ATCC medium 2154.
Screw cap on tightly and incubate at a 15° horizontal slant at 35°C. Observe the culture daily and transfer when many trophozoites are observed.
Name of Depositor LS Diamond
Chain of Custody
ATCC <-- LS Diamond <-- JE Tobie
Year of Origin 1949
References 参考文献
Adagu IS, et al. In vitro activity of nitazoxanide and related compounds against isolates of Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. J. Antimicrob. Chemother. 49: 103-111, 2002. PubMed: 11751773
Beaulieu BB Jr., et al. Metronidazole metabolism in cultures of Entamoeba histolytica and Trichomonas vaginalis. Antimicrob. Agents Chemother. 20: 410-414, 1981. PubMed: 6272633
Clark CG, Diamond LS. Intraspecific variation and phylogenetic relationships in the genus Entamoeba as revealed by riboprinting. J. Eukaryot. Microbiol. 44: 142-154, 1997. PubMed: 9109261
Diamond LS. Techniques of axenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae. J. Parasitol. 54: 1047-1056, 1968. PubMed: 4319346
Gillin FD, Diamond LS. Attachment of Entamoeba histolytica to glass in a defined maintenance medium: Specific requirement for cysteine and ascorbic acid. J. Protozool. 27: 474-478, 1980. PubMed: 6260930
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
Gonzalez-Ruiz A, et al. Diagnosis of amebic dysentery by detection of Entamoeba histolytica fecal antigen by an invasive strain-specific, monoclonal antibody-based enzyme-linked immunosorbent assay. J. Clin. Microbiol. 32: 964-970, 1994. PubMed: 8027351
Simione FP Jr., et al. The use of plastic ampoules for freeze preservation of microorganisms. Cryobiology 14: 500-502, 1977. PubMed: 891238
相关产品
嗜盐盐单胞菌 Halomonas halophila
Xenorhabdus beddingii
Halogranum salarium
Thermogladius calderae
Methanosarcina subterranea
Halohasta litchfieldiae
Halorubrum lipolyticum
Geoglobus acetivorans
Pyrobaculum calidifontis
Methanobacterium arcticum
Pedococcus cremeus
Pedococcus dokdonensis
Pelagibacterium luteolum
Clostridium innocuum
Clostridium indicum
关注
拨打电话
留言咨询