Chapter 19
Coelenterata
Subprotocol 1
Interstitial Cells from Hydra (Hydroidea)
Materials
- Microscope
- Centrifuge and centrifuge tube
- 125-ml culture flasks
- Leighton tubes
- Hemocytometer
- Hydra culture solution (see Appendix 1)
- Hydra growth medium (see Appendix 1)
- Neomycin
- Streptomycin
- Fumidil B (WoodsEnd, Inc)
- Penicillin
- Cultured Hydra littoralis (see Comments)
- Gauze
Procedure
Hydra seem to have intracellular parasitic protozoa. It is essential to remove these microorganisms. The following is the protocol ofYu-Ying et
a1. (1963) to prevent infection by these microorganisms.
PROTOCOL
Equipment
Solutions
Antibiotics
Animals
Miscellaneous
1. Culture hydra for 2 days without feeding in sterilized culture solution Elimination of
containing 0.125 g neomycin, 0.125 g streptomycin, 0.200 g Fumidil B, intracellular
and 500 IU penicillin per 1.
protozoa
1. Trypsinize the hydra tissues according to the methods of Youngner Dissociation
(1954; see Comments).
2. Filter the cell suspension through four layers of gauze.
3. Centrifuge the filtrate at 50 g for 10 min to remove nematocysts.