80
Methods for Setting Up Primary Cultures Specific to Animal Groups
Fig. 6. Migrated cells from explants (EX) of Drosophila imaginal discs. x 80 From Ui, K., et ai.,
"Cell lines from imaginal discs of
Drosophila melanogaster", In Vitro
Cellular and Developmental Biology-Animal, Voi.23 pp. 708. Copyright © 1987 by the Society for In
Vitro Biology (formerly the Tissue Culture Association). Reproduced with permission of the
copyright owner
Fig. 7. Spherical vesicles (arrows) formed
on the fragments of Drosophila imaginal
discs. x 80. From Ui, K., et ai., ditto
D. melanogaster cell lines from imaginal discs have been said to differentiate into cuticular structures when implanted into metamorphosing
larvae (Ui et aI., 1987) .
• 111 Comments
Drosophila Wyss (1982) used his ZW medium (see Appendix 1) by fortifying it with
extract an extract of adult D. melanogaster and insulin. The method to prepare
the fly extract is as follows:
1. Accumulate 100 g flies in a freezer at -70°C.
2. Add 500 ml cold 0.2 M ammonium acetate in water, and macerate in a
kitchen grinder at top speed for 1 min.
3. Centrifuge the homogenate at 9,000 g for 30 min at 4°C.
4. Filter the supernatant through glass wool.
5. Add an equal volume of diethyl ether to the filtrate.
6. Centrifuge the turbid water layer at 9,000 g for 30 min at 4°C.
7. Remove the ether and the leathery interphase.
8. Evaporate the remaining ether.
9. Heat the water phase to 60°C for 10 min. This treatment produces massive precipitation.
Methods for Setting Up Primary Cultures Specific to Animal Groups
Fig. 6. Migrated cells from explants (EX) of Drosophila imaginal discs. x 80 From Ui, K., et ai.,
"Cell lines from imaginal discs of
Drosophila melanogaster", In Vitro
Cellular and Developmental Biology-Animal, Voi.23 pp. 708. Copyright © 1987 by the Society for In
Vitro Biology (formerly the Tissue Culture Association). Reproduced with permission of the
copyright owner
Fig. 7. Spherical vesicles (arrows) formed
on the fragments of Drosophila imaginal
discs. x 80. From Ui, K., et ai., ditto
D. melanogaster cell lines from imaginal discs have been said to differentiate into cuticular structures when implanted into metamorphosing
larvae (Ui et aI., 1987) .
• 111 Comments
Drosophila Wyss (1982) used his ZW medium (see Appendix 1) by fortifying it with
extract an extract of adult D. melanogaster and insulin. The method to prepare
the fly extract is as follows:
1. Accumulate 100 g flies in a freezer at -70°C.
2. Add 500 ml cold 0.2 M ammonium acetate in water, and macerate in a
kitchen grinder at top speed for 1 min.
3. Centrifuge the homogenate at 9,000 g for 30 min at 4°C.
4. Filter the supernatant through glass wool.
5. Add an equal volume of diethyl ether to the filtrate.
6. Centrifuge the turbid water layer at 9,000 g for 30 min at 4°C.
7. Remove the ether and the leathery interphase.
8. Evaporate the remaining ether.
9. Heat the water phase to 60°C for 10 min. This treatment produces massive precipitation.