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Methods for Setting Up Primary Cultures Specific to Animal Groups
Culture 1. Cut the ovaries into small fragments.
set-up 2. Transfer the fragments, with the culture medium, into a culture vessel.
3. Keep the culture at 30°C.
Results
According to Giauffret et al. (1968), the ovarioles from larvae produce
many different types of cells (Fig. 1). At the beginning of culture, elongated fibroblast-like cells appear. These cells are inclined to aggregate to
form networks or bundles. Later, larger and more elongated cells appear
and form a monolayer of cells. These cells survive for many weeks and
show mitosis during that period.
Subprotocol 2
Embryos
Sohi and Ennis (1981) obtained a continuous cell line from the embryos
of the sawfly Neodiprion lecontei using the following method.
Materials
Equipment - Dissecting microscope
- Scissors
- Forceps
- Needles
- Small petri dishes
Solutions - Quadramine X-IOO (Soden Chemicals)
- Locke's solution (see Appendix 1)
Animals - Eggs of the sawfly
Procedure
Sterilization 1. Surface sterilize the eggs of the sawfly by soaking them in a 1 % aqueous solution of Quadramine X-IOO for 1 min.
2. Rinse the eggs with three changes of Locke's saline, and place them in
a small petri dish with the solution.
Dissection 1. Cut the egg chorion and remove the embryos individually with fine
forceps.
2. Transfer the embryos into a dish containing culture medium.
3. Remove the yolks attached to the embryos.
Methods for Setting Up Primary Cultures Specific to Animal Groups
Culture 1. Cut the ovaries into small fragments.
set-up 2. Transfer the fragments, with the culture medium, into a culture vessel.
3. Keep the culture at 30°C.
Results
According to Giauffret et al. (1968), the ovarioles from larvae produce
many different types of cells (Fig. 1). At the beginning of culture, elongated fibroblast-like cells appear. These cells are inclined to aggregate to
form networks or bundles. Later, larger and more elongated cells appear
and form a monolayer of cells. These cells survive for many weeks and
show mitosis during that period.
Subprotocol 2
Embryos
Sohi and Ennis (1981) obtained a continuous cell line from the embryos
of the sawfly Neodiprion lecontei using the following method.
Materials
Equipment - Dissecting microscope
- Scissors
- Forceps
- Needles
- Small petri dishes
Solutions - Quadramine X-IOO (Soden Chemicals)
- Locke's solution (see Appendix 1)
Animals - Eggs of the sawfly
Procedure
Sterilization 1. Surface sterilize the eggs of the sawfly by soaking them in a 1 % aqueous solution of Quadramine X-IOO for 1 min.
2. Rinse the eggs with three changes of Locke's saline, and place them in
a small petri dish with the solution.
Dissection 1. Cut the egg chorion and remove the embryos individually with fine
forceps.
2. Transfer the embryos into a dish containing culture medium.
3. Remove the yolks attached to the embryos.