18 Platyhelminthes
213
Results
According to the results of the study of Dieckmann -Shuppert et al. (1989),
the hybridoma cells remained viable for at least 10 weeks. The cells formed
colonies 2 weeks after fusion, and cell multiplication continued for 6
weeks. The hybridoma cells showed E. multilocularis antigens on their
surface.
Subprotocol 3
Cells from Schistosoma mansoni (Trematoda)
Materials
- CO 2 incubator
- Leighton tube
- Pasteur pipette
- Petri dish
- Tube (8.5 x 1.5 cm)
- Long-handled sharp scissors
- Plastic disposable pipette
- Beakers
- Medium 1779 for Schistosoma (see Appendix 1)
- Schistosoma salt solution (see Appendix 1)
- Schistosoma mansoni (obtained from mice infected for 4 weeks)
Procedure
1. Collect the worms from the blood of infected mice by suction with a
Pasteur pipette into a beaker.
2. Transfer the worms to a petri dish.
3. Wash the worms several times with Schistosoma salt solution, and remove host tissues if present.
4. Cool for 10-15 min at 5°C.
5. Transfer the worms to a tube.
6. Wash once with Schistosoma salt solution containing 0.25% trypsin.
Equipment
Solutions
Animals
Separation
of worms
1. Mince the worms with long-handled sharp pointed scissors in 0.25 ml Dissection
trypsin solution.
2. Increase the volume to 3.0 ml by adding trypsin solution.
3. Trypsinize the fragments for 10 min at room temperature. Change the
supernatant solution once or twice during trypsinization.
4. Remove the trypsin solution with a Pasteur pipette.
5. Wash the fragments three times in 3.0 ml medium 1779.