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Product Name:
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HM-1:IMSS
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Manufacturer:
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BEI Resources
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Taxonomy:
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Protozoa Classification: Entamoebidae, Entamoeba
Species: Entamoeba histolytica
Strain: HM-1:IMSS
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Additional Information:
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E. histolytica is a pathogenic protozoan parasite and causative agent of amebiasis, an intestinal infection that predominantly infects humans and other primates in developing countries, with symptoms ranging from asymptomatic colonization to extraintestinal, disseminated disease.6,7,8 The E. histolytica life cycle consists of a highly resistant environmental cyst with a protective, chitin-rich cell wall and a dividing trophozoite, which establishes infection through excystation in the colon.7,9 Infection occurs through shedding of cysts in feces and the ingestion of cysts via contaminated water and vegetables.9 E. histolytica has been shown to cause host tissue damage through amoebic trogocytosis in a mouse model.10
Derived from ATCC® 30459™
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Material Provided:
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Each vial contains approximately 0.5 mL of cells in cryopreservative. Please refer to the Certificate of Analysis for the specific culture media used for each lot and refer to Appendix I for cryopreservation instructions.
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Packing/Storage:
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NR-178 was packaged aseptically in cryovials and is provided frozen on dry ice. The product should be stored at -130°C or colder, preferably in the vapor phase of a liquid nitrogen freezer. If liquid nitrogen storage facilities are not available, frozen cryovials may be stored at 70°C or colder for approximately one week.
Note: Do not under any circumstances store vials at temperatures warmer than -70°C. Storage under these conditions will result in the death of the culture.
To ensure the highest level of viability, the culture should be initiated immediately upon receipt. Any warming of the product during shipping and transfer must be avoided, as this will adversely affect the viability of the product. For transfer between freezers and for shipping, the product may be placed on dry ice for brief periods, although use of a portable liquid nitrogen carrier is preferred. Please read the following recommendations prior to using this material.
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Growth Conditions:
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Growth Media:
Liver Digest – Yeast Extract – Iron (LYI) Entamoeba medium supplemented with 10% heat-inactivated adult bovine serum (HIBS) or equivalent (Appendix II)
Incubation:
Temperature: 35°C to 37°C
Atmosphere: Axenic and microaerophilic
Propagation:
1. To establish a culture from the frozen state, place a vial in a 35°C water bath for 2 to 3 minutes, until thawed. Immerse the vial just enough to cover the frozen material. Do not agitate the vial.
2. Transfer the vial contents to a 16 × 125 mm screw-capped borosilicate glass test tube containing 13 mL of growth medium.
3. Screw the cap on tightly and incubate at a 15° horizontal slant at 35°C to 37°C. Observe the culture daily and subculture when peak trophozoite density is observed.
Maintenance:
1. When the culture is at or near peak density, ice the culture for 10 minutes and gently invert 20 times.
2. Add 12 mL of freshly prepared growth media to two sterile tubes.
3. Aseptically transfer a 0.1 mL and 0.25 mL aliquot of E. histolytica, strain HM-1:IMSS to the tubes in step 2.
4. Screw the cap on tightly and incubate at a 15° horizontal slant at 35°C to 37°C. Observe the culture daily and subculture when peak trophozoite density is observed.
Please refer to Appendix I for cryopreservation instructions.
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Disclaimers:
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You are authorized to use this product for research use only. It is not intended for human use. Use of this product is subject to the terms and conditions of the BEI Resources Material Transfer Agreement (MTA). The MTA is available on our Web site at www.beiresources.org. While BEI Resources uses reasonable efforts to include accurate and up-to-date information on this product sheet, neither ATCC® nor the U.S. Government makes any warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. Neither ATCC® nor the U.S. Government warrants that such information has been confirmed to be accurate. This product is sent with the condition that you are responsible for its safe storage, handling, use and disposal. ATCC® and the U.S. Government are not liable for any damages or injuries arising from receipt and/ or use of this product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, the U.S. Government, ATCC®, their suppliers and contributors to BEI Resources are not liable for damages arising from the misidentification or misrepresentation of products. |
References:
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1. Landa, L., B. Sepúlveda, and M. De la Torre. “Advances in Methods of Entamoeba histolytica Culture.” Arch. Invest. Med. (Mex.) 1 (1970): 9-14. PubMed: 4332884.
2. Diamond, L. S. “Techniques of Axenic Cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-Like Amebae.” J. Parasitol. 54 (1968): 1047-1056. PubMed: 4319346.
3. Diamond, L. S. “Axenic Cultivation of Entamoeba histolytica.” Science 134 (1961): 336-337. PubMed: 13722605.
4. Clark, C. G. and L. S. Diamond. “Methods for Cultivation of Luminal Parasitic Protists of Clinical Importance.” Clin. Microbiol. Rev. 15 (2002): 329–341. PubMed: 12097242.
5. Loftus, B. J. and N. Hall. “The Genome of the Protist Parasite Entamoeba histolytica.” Nature 433 (2005): 865-868. PubMed: 15729342.
6. Pritt, B. S. and C. G. Clark. “Amebiasis.” Mayo Clin. Proc. 83 (2008): 1154-1159. PubMed: 18828976.
7. Parija, S. C., J. Mandal and D. K. Ponnambath. “Laboratory Methods of Identification of Entamoeba histolytica and Its Differentiation from Look-Alike Entamoeba spp.” Trop. Parasitol. 4 (2014): 90-95. PubMed: 25250228.
8. Ngui, R., et al. “Differentiating Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii Using Nested Polymerase Chain Reaction (PCR) in Rural Communities in Malaysia.” Parasit. Vectors 5 (2012): 187. PubMed: 22947430.
9. Marie, C. and W. A. Petri Jr. “Regulation of Virulence of E. histolytica.” Annu. Rev. Microbiol. 68 (2014): 493-520. PubMed: 25002094.
10. Ralston, K. S. ”Taking a Bite: Amoebic Trogocytosis in Entamoeba histolytica and Beyond.” Current Opin. Microbiol. 28 (2015): 26-35. PubMed: 26277085.
11. Stanley, S. L., Jr. “The Entamoeba histolytica Genome: Something Old, Something New, Something Borrowed and Sex Too?” Trends Parasitol. 21 (2005): 451-453. PubMed: 16098811.
12. Loftus, B. J. and N. Hall. “Entamoeba: Still More to be Learned from the Genome.” Trends Parasitol. 21 (2005): 453. PubMed: 16099723.
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Citation:
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Acknowledgment for publications should read “The following reagent was obtained through BEI Resources, NIAID, NIH: Entamoeba histolytica, Strain HM-1:IMSS, NR-178.”
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Biosafety Level:
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2
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories (BMBL). Current Edition. Washington, DC: U.S. Government Printing Office.
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