66
Methods for Setting Up Primary Cultures Specific to Animal Groups
5. Transfer the fragments, with the culture medium, into culture vessels.
6. Culture at 25°C.
Results
Cell migration may occur from fragmented ganglia (Fig. 19). Sometimes
the migrated cells proliferate. These proliferating cells are usually glial
cells. Nerve cells and neurosecretory cells do not multiply.
Subprotocol12
Embryos
Embryos consist of quickly growing cells. There are many actively proliferating cells in embryos. Therefore, it is considered that this developmental stage provides proliferating cells even in vitro.
Materials
Equipment The equipment used is the same as given for subprotocoll.
Solutions - Physiological solution
- Culture media
Animals - Eggs oflepidopteran insects
Procedure
Sterilization 1. Sterilize the surface of the eggs or egg masses by submersion in 70%
ethanol for 3-10 min.
2. Wash the eggs in sterile distilled water.
3. Place the eggs in physiological solution on a Maximov slide, or in a
small petri dish.
Dissection 1. Cut or tear the chorion of the eggs with a sharp knife, needles, or forceps.
2. Squeeze the embryos and yolk from the eggs.
3. Discard the chorion.
Culture l. Remove the yolk enveloping the embryos, or, when the embryos are
set-up
matured, cut the embryos and remove the yolk, which is taken into
the abdomen of the embryos by dorsal closure.
2. Collect the embryos, and transfer them to fresh physiological solution.
Methods for Setting Up Primary Cultures Specific to Animal Groups
5. Transfer the fragments, with the culture medium, into culture vessels.
6. Culture at 25°C.
Results
Cell migration may occur from fragmented ganglia (Fig. 19). Sometimes
the migrated cells proliferate. These proliferating cells are usually glial
cells. Nerve cells and neurosecretory cells do not multiply.
Subprotocol12
Embryos
Embryos consist of quickly growing cells. There are many actively proliferating cells in embryos. Therefore, it is considered that this developmental stage provides proliferating cells even in vitro.
Materials
Equipment The equipment used is the same as given for subprotocoll.
Solutions - Physiological solution
- Culture media
Animals - Eggs oflepidopteran insects
Procedure
Sterilization 1. Sterilize the surface of the eggs or egg masses by submersion in 70%
ethanol for 3-10 min.
2. Wash the eggs in sterile distilled water.
3. Place the eggs in physiological solution on a Maximov slide, or in a
small petri dish.
Dissection 1. Cut or tear the chorion of the eggs with a sharp knife, needles, or forceps.
2. Squeeze the embryos and yolk from the eggs.
3. Discard the chorion.
Culture l. Remove the yolk enveloping the embryos, or, when the embryos are
set-up
matured, cut the embryos and remove the yolk, which is taken into
the abdomen of the embryos by dorsal closure.
2. Collect the embryos, and transfer them to fresh physiological solution.