18 Platyhelminthes
217
- Culture medium (Locke's solution diluted to between 1/10 and 1/12 Solutions
with boiled well water, with 0.1-1.0 % glucose and 25% sheep serum
added)
- Planaria Planaria dorotocephala
Animals
Procedure
1. Surface sterilize animals by irradiation from a UV lamp, as described Sterilization
for the sterilization process in subprotocol 4.
1. Cut the animal into small pieces, 0.3-0.5 mm diameter, which are ap- Cutting
proximately between 1/400 and 1/120 the volume of an animal of standard dimensions. Such small pieces, if placed in water instead of in
culture media, do not regenerate, but will die within a few hours.
1. Using a pipette, transfer the tissue fragments onto a cover glass with Culture
culture medium.
set-up
2. Invert the cover glass over a depression slide, in the bottom of which a
small drop of water had been placed.
3. Seal the two together with vaseline or mixture of vaseline and paraffin.
4. Culture at 20°C or lower.
Results
Cells migrated and survived for 15 days. The migrated cells showed
pseudopodial activity, and amitotic division. The migrated cells did not
form cell sheets. The addition of a mixture of glucose and peptone perceptibly stimulated the cells to migrate and divide.
Subprotocol 6
Neoblasts from P/anaria (Turbellaria)
The following is the method of Betchaku (1967) for the culture of
neoblasts isolated from Planaria.
Materials
- Dissecting microscope
- Inverted microscope
- Scissors
- Forceps
- Needle
- Pasteur pipettes
Equipment
217
- Culture medium (Locke's solution diluted to between 1/10 and 1/12 Solutions
with boiled well water, with 0.1-1.0 % glucose and 25% sheep serum
added)
- Planaria Planaria dorotocephala
Animals
Procedure
1. Surface sterilize animals by irradiation from a UV lamp, as described Sterilization
for the sterilization process in subprotocol 4.
1. Cut the animal into small pieces, 0.3-0.5 mm diameter, which are ap- Cutting
proximately between 1/400 and 1/120 the volume of an animal of standard dimensions. Such small pieces, if placed in water instead of in
culture media, do not regenerate, but will die within a few hours.
1. Using a pipette, transfer the tissue fragments onto a cover glass with Culture
culture medium.
set-up
2. Invert the cover glass over a depression slide, in the bottom of which a
small drop of water had been placed.
3. Seal the two together with vaseline or mixture of vaseline and paraffin.
4. Culture at 20°C or lower.
Results
Cells migrated and survived for 15 days. The migrated cells showed
pseudopodial activity, and amitotic division. The migrated cells did not
form cell sheets. The addition of a mixture of glucose and peptone perceptibly stimulated the cells to migrate and divide.
Subprotocol 6
Neoblasts from P/anaria (Turbellaria)
The following is the method of Betchaku (1967) for the culture of
neoblasts isolated from Planaria.
Materials
- Dissecting microscope
- Inverted microscope
- Scissors
- Forceps
- Needle
- Pasteur pipettes
Equipment