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Organ Culture
Procedure
The following is the method of Freeman and Costlow (1979) for the cultivation of mantle tissues from a barnacle.
Dissection 1. Extirpate the prosoma from the animal (cf. chapter 12, Fig.14).
2. Excise the inner mantle tissue.
3. Rinse the tissue briefly in fIltered seawater.
Culture 1. Cut the tissue to make explants smaller than 3 mm in any dimension.
set-up 2. Place the explants in 0.5 ml culture medium in a 100 x 13-mm test
tube and place the test tube at an angle of approximately 30° from
horizontal in a test tube rack. The rack is then placed on a shaker,
which oscillates at a rate of 80 oscillations/min.
Results
According to Freeman and Costlow (1979), apolysis and cell enlargement were completed in the cultured inner mantle tissues.
Subprotocol 4
x- and V-Organs of Crustaceans (Crustacea)
Materials
Equipment - Recirculating seawater system
- Scissors
- Forceps
- Dissecting tray
- Pasteur pipettes
- Double-well organ culture dish (no. 3010; Falcon; Becton-Dickinson)
- 02-supply system (optional)
Solutions - Seawater
- Culture medium (TC-199 medium supplemented with 5% FBS and
20 mg/ml BSA (Sigma), or L-15 medium mixed with an equal volume
of crab saline; see Appendix 1)
Animals - Crabs (Pachygrapsus crassipes or Cardisoma carnifex) or lobsters
(Panulirus marginatus)
Miscellaneous - Ice
Procedure
Maintenance 1. Place the animals into aquariums equipped with a recirculating seaof animals
water system at lrC and feed the animals on pieces of smelt.
Organ Culture
Procedure
The following is the method of Freeman and Costlow (1979) for the cultivation of mantle tissues from a barnacle.
Dissection 1. Extirpate the prosoma from the animal (cf. chapter 12, Fig.14).
2. Excise the inner mantle tissue.
3. Rinse the tissue briefly in fIltered seawater.
Culture 1. Cut the tissue to make explants smaller than 3 mm in any dimension.
set-up 2. Place the explants in 0.5 ml culture medium in a 100 x 13-mm test
tube and place the test tube at an angle of approximately 30° from
horizontal in a test tube rack. The rack is then placed on a shaker,
which oscillates at a rate of 80 oscillations/min.
Results
According to Freeman and Costlow (1979), apolysis and cell enlargement were completed in the cultured inner mantle tissues.
Subprotocol 4
x- and V-Organs of Crustaceans (Crustacea)
Materials
Equipment - Recirculating seawater system
- Scissors
- Forceps
- Dissecting tray
- Pasteur pipettes
- Double-well organ culture dish (no. 3010; Falcon; Becton-Dickinson)
- 02-supply system (optional)
Solutions - Seawater
- Culture medium (TC-199 medium supplemented with 5% FBS and
20 mg/ml BSA (Sigma), or L-15 medium mixed with an equal volume
of crab saline; see Appendix 1)
Animals - Crabs (Pachygrapsus crassipes or Cardisoma carnifex) or lobsters
(Panulirus marginatus)
Miscellaneous - Ice
Procedure
Maintenance 1. Place the animals into aquariums equipped with a recirculating seaof animals
water system at lrC and feed the animals on pieces of smelt.