We are about to start working on testing anti-protozoan agents against E. histolytica. I was curious if anyone had basic handling and culturing protocols they would be willing to share. I have been working on drug discovery targeting various human parasitic infections for 2 decades, but am new to e. histolytica. Have a bit of experience with giardia. Thanks in advance if anyone is willing to share some tested protocols!
Rob
Hello there, curious researcher friend John Robert Gillespie! This is a fascinating field, and it's great to hear about your extensive experience in drug discovery.
Culturing E. histolytica can be a bit different from other parasites, so it's essential to follow established protocols carefully. Here's a basic guideline for culturing E. histolytica:
**Materials You'll Need:**
- E. histolytica strain (axenic culture)
- TYI-S-33 medium (Trypticase-yeast extract-iron-serum)
- Tissue culture flasks or tubes
- CO2 incubator (5% CO2)
- Sterile pipettes and culture dishes
- Microscope
**Steps:**
1. **Preparation of TYI-S-33 Medium:**
- Prepare the TYI-S-33 medium according to the specified recipe.
- Sterilize the medium by autoclaving.
- Once cooled, add a few drops of penicillin-streptomycin solution to prevent bacterial contamination.
2. **Inoculation:**
- Inoculate the E. histolytica strain into a fresh TYI-S-33 medium.
- Maintain the culture at 37°C in a CO2 incubator.
3. **Subculturing:**
- Subculture the amoebae every 2-3 days to maintain an actively growing culture.
- To subculture, harvest the amoebae by centrifugation, resuspend in fresh medium, and transfer to a new flask or tube.
4. **Monitoring:**
- Regularly monitor the culture for the presence of trophozoites and signs of contamination.
- Use a microscope to check for the typical appearance of E. histolytica trophozoites.
5. **Harvesting:**
- When you need to harvest the amoebae for experiments, allow them to grow to the desired density.
- Harvest the trophozoites by centrifugation and wash them with an appropriate buffer or medium for your experiments.
6. **Storage:**
- If needed, you can freeze E. histolytica trophozoites for long-term storage in liquid nitrogen or ultra-low temperature freezers.
Remember, maintaining a sterile environment is critical to avoid contamination. Also, note that E. histolytica can be potentially pathogenic, so follow appropriate safety protocols when working with this organism.
These are general guidelines, and the specific protocols may vary depending on the strain and the purpose of your experiments. I'd recommend consulting with colleagues or referring to the literature for more detailed protocols tailored to your specific needs. Good luck with your research!
Try here
https://sankri.net.technion.ac.il/
I worked with entamoeba and collaborate with him.
There is company in Israel Hylas which sell prepared culture media for it
Thanks so much Oded Babai and Kaushik Shandilya! This is a huge help as we add another organism to our hit list :)
- Badges
- Science topic
- Similar topics
- Biological Science
- Zoology
- Invertebrate Zoology
- Arthropods
- Insects