Chapter 12
Arthropods Other Than Insects
Subprotocol 1
Tick (Arachnida) Embryos
Arachnida is the second most studied arthropoda for cell culture. Several
continuously growing cell lines have been established from tick embryos.
Materials
- Centrifuge and centrifuge tube
- Petri dish (90 mm diameter)
- Glass rod
- Sintered glass funnel (pore size 100-200 jlm)
- Rubber policeman
- Culture vessel
- Pasteur pipettes
PROTOCOL
Equipment
- Leibovitz's L-IS medium (see Appendix 1)
Solutions
- Antibiotic mixture (100 IV/ml penicillin G sodium salt and 1 mg/ml
streptomycin sulfate)
- Dulbecco's phosphate-buffered saline A (PBSA; Oxoid)
- Engorged female ticks
Animals
- Roccal (Winthrop Laboratories)
Others
- Filter paper (heat sterilized)
Procedure
1. Wash engorged female ticks after completion of blood meal and before they have laid any eggs.
2. Shake the ticks in two changes of 1 : 10 Roccal containing benzalkonium chloride, and then leave the ticks to soak in a small container
with 1 : 10 Roccal for 20 min.
3. Rinse the ticks quickly in ethanol, and then twice in a sterile solution
of the antibiotic mixture.
4. Dry the ticks on sterile filter paper, and put them in a sterile petri dish
in a moist chamber.
S. Incubate the ticks at 28°C for oviposition. Oviposition will start 1-2
days after the beginning of the incubation.
Aseptic
oviposition
Arthropods Other Than Insects
Subprotocol 1
Tick (Arachnida) Embryos
Arachnida is the second most studied arthropoda for cell culture. Several
continuously growing cell lines have been established from tick embryos.
Materials
- Centrifuge and centrifuge tube
- Petri dish (90 mm diameter)
- Glass rod
- Sintered glass funnel (pore size 100-200 jlm)
- Rubber policeman
- Culture vessel
- Pasteur pipettes
PROTOCOL
Equipment
- Leibovitz's L-IS medium (see Appendix 1)
Solutions
- Antibiotic mixture (100 IV/ml penicillin G sodium salt and 1 mg/ml
streptomycin sulfate)
- Dulbecco's phosphate-buffered saline A (PBSA; Oxoid)
- Engorged female ticks
Animals
- Roccal (Winthrop Laboratories)
Others
- Filter paper (heat sterilized)
Procedure
1. Wash engorged female ticks after completion of blood meal and before they have laid any eggs.
2. Shake the ticks in two changes of 1 : 10 Roccal containing benzalkonium chloride, and then leave the ticks to soak in a small container
with 1 : 10 Roccal for 20 min.
3. Rinse the ticks quickly in ethanol, and then twice in a sterile solution
of the antibiotic mixture.
4. Dry the ticks on sterile filter paper, and put them in a sterile petri dish
in a moist chamber.
S. Incubate the ticks at 28°C for oviposition. Oviposition will start 1-2
days after the beginning of the incubation.
Aseptic
oviposition