288
Organ Culture
Subprotocol 2
Integuments of Earthworms {Oligochaeta}
Materials
Equipment - The equipment used is the same as given for subprotocoll
Solutions - Wolff et al.'s medium C (see Appendix I)
Animals - Earthworms E. foetida
Procedure
Follow steps 1 and 2 of subprotocoll for sterilization.
Anesthesia 1. Anesthetize the worms by chilling them at -lO°C.
Dissection I. Cut integuments, with connected muscle tissues, of three to four segments from the dorsal part of the worms.
2. Cut the integuments into three pieces.
Culture I. Culture the integument fragments in Wolff et aI:s medium C at 23°C.
set-up
Results
According to Berjon (1965), over a period of 5 or 7 days culture, mucous
glands in the integument decreased their volume and granules in granular cells disappeared. However, if the brains of mature worms were
co cultured with the integument, these cells maintained their appearance
similar to that seen in vivo.
Subprotocol 3
Various Organs of Leeches (Hirudinea)
Materials
Equipment - The equipment used is the same as given for subprotocoll
Solutions - Durchon and Schaller's medium (see Appendix 1)
Animals - Leeches Hirudo medicinalis
Procedure
Sterilization 1. Keep the leeches in sterile water containing penicillin G (500 IU/ml)
and streptomycin (0.025 mg/ml).
2. Sterilize the leeches by UV radiation.