29 Coelenterata
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1. Cut the base of the stalk and the upper part of the body bearing the Dissection
tentacles and gonophores using a sterile knife or scissors.
2. Wash the separated stalks several times in autoclaved seawater.
3. Cut the stalks on a sterile dish into 1.0-mm lengths.
1. Using a Pasteur pipette, transfer the fragments to hanging drops of Culture
autoclaved seawater containing penicillin (100 IU/ml).
set-up
2. Maintain the culture at 17°_22°C.
Results
According to Rudzinska (1951), stalk fragments that were 0.5 or 2.0 mm
in length gave unsatisfactory results. Migration of endodermal cells
started at both cut edges of the fragment after 16 h. The migrating cells
became oblong and narrow. These cells formed long projections. The
ectodermal cells showed a tendency to migrate in sheets composed of
cells with small projections along the free borders of the sheets. Such
cultures were maintained in hanging drops for over 2 weeks. It was found
that filter-sterilized seawater or White's M6W medium were unsuitable
for use in culture. Cultures of hydra stalks were also attempted in roller
tubes or Carrel flasks. These containers proved unsuitable for the culture of regenerating hydra fragments.
Subprotocol 3
Gorgonian Tissues of Hydra (Hydrozoa)
Materials
- Membrane filter system
- Culture vessels
- Pasteur pipettes
- Seawater
- Hydra Eunicella stricta or Lophogorgia sarmentosa
Procedure
Culture of gorgonian tissues can be undertaken in non-sterile conditions
by using seawater filtered through a 0.45-flm pore size membrane filter
as the medium.
1. Excise gorgonian tissues from the animal.
2. Maintain the tissues in the seawater medium.
3. Keep the culture at 10°-15°C.
Equipment
Solutions
Animals
Culture
set-up