12 Arthropods Other than Insects
123
Fig. 1. Cell migration in primary culture,
28 days after the initiation of the culture,
from attached cell clumps of the embryo
of the tick Boophilus microplus. x 75. From
Pudney, M. et ai., 1973, with permission
vessel, subculture may be possible (see Comments). By repeating subcultures, Pudney et al. (1973) and Varma et al. (1975) obtained several
continuous cell lines from embryos of Boophilus microplus and Rhipicephalus appendiculatus, respectively. Rahman (1981) attempted to culture embryonic cells of Hyalomma rufipes using a similar procedure,
and obtained fibroblast-like cells that survived for 2-3 weeks.
Subprotocol 2
Crayfish (Crustacea) Embryos
•
Materials
- Centrifuge and centrifuge tubes
- Culture flasks (30 ml; Falcon, Becton-Dickinson)
- Forceps
- Sharp knife
- Pasteur pipettes
- Erlenmeyer flask
- Magnetic stirrer and magnetic bar
- Rinaldini's salt solution (see Appendix 1)
- Culture medium
- Crayfish Astacus pallipes and Astacus leptodactylus
• • Procedure
The following procedure is based on the methods of Quiot and Vey (1978).
Equipment
Solutions
Animals
1. Remove 30-35 eggs from female crayfish when the eyes of the em- Sterilization
bryos become visible through the chorionic membrane.
2. Surface sterilize the eggs in a hypochlorite solution for 5 min.
3. Wash the eggs in sterilized distilled water.
123
Fig. 1. Cell migration in primary culture,
28 days after the initiation of the culture,
from attached cell clumps of the embryo
of the tick Boophilus microplus. x 75. From
Pudney, M. et ai., 1973, with permission
vessel, subculture may be possible (see Comments). By repeating subcultures, Pudney et al. (1973) and Varma et al. (1975) obtained several
continuous cell lines from embryos of Boophilus microplus and Rhipicephalus appendiculatus, respectively. Rahman (1981) attempted to culture embryonic cells of Hyalomma rufipes using a similar procedure,
and obtained fibroblast-like cells that survived for 2-3 weeks.
Subprotocol 2
Crayfish (Crustacea) Embryos
•
Materials
- Centrifuge and centrifuge tubes
- Culture flasks (30 ml; Falcon, Becton-Dickinson)
- Forceps
- Sharp knife
- Pasteur pipettes
- Erlenmeyer flask
- Magnetic stirrer and magnetic bar
- Rinaldini's salt solution (see Appendix 1)
- Culture medium
- Crayfish Astacus pallipes and Astacus leptodactylus
• • Procedure
The following procedure is based on the methods of Quiot and Vey (1978).
Equipment
Solutions
Animals
1. Remove 30-35 eggs from female crayfish when the eyes of the em- Sterilization
bryos become visible through the chorionic membrane.
2. Surface sterilize the eggs in a hypochlorite solution for 5 min.
3. Wash the eggs in sterilized distilled water.