262
Organ Culture
- Remove the two mesothoracic legs of fifth ins tar nymphs of the cockroach Leucophaea maderae at the trochanter-femoral joint 2 days after molting
- Open the stumps from 10 to 21 days after removal of the legs, and
dissect out the regenerating tissues
Results
According to Marks and Reinecke (1964), in a culture of regenerating
tissues of the legs of L. maderae, active cell migration, pinocytosis, and
phagocytosis of cells in the explants were observed. In addition, the production of vesicles by explanted tissues was seen. These vesicles were
hollow, spherical structures that, when mature, consisted of a single layer
of thin epithelial cells, which became interlaced with attenuated fiberlike cells. Later, Marks and Leopold (1970) obtained the chitinous cuticle
on the explants. However, the cuticle formed in vitro remained thin and
only partially developed. The heavy layer of endocuticle was largely absent. Bulliere and Bulliere (1977) cultured the regenerating appendage
tissues of embryonic legs of Blaberus craniifer in a salt mixture (NaCI
12.65 g, KCI l.61 g, and CaCl 2 l.26 g in 1,000 ml H 2 0) containing 25%
bovine serum. The embryo of this cockroach has two molting cycles,
and regeneration of appendage is possible only during the first molting
cycle. When tarsus segments of the embryonic appendage were removed
and the appendage was cultured, regeneration took place inside the cuticle of the stump. The addition of molting hormone to the medium inhibited regeneration.
Subprotocol 19
Tumor Tissues of DrosophilQ (Diptera)
Materials
Equipment - Dissecting microscope
- Maximov slide
- Forceps
- Needles
- Pasteur pipettes
- Culture vessel
Solutions - Ringer's solution
- Culture medium (Kuroda and Tamura's medium; see Appendix 1)
Animals - Aseptically reared larvae of D. melanogaster (preferably strain v tu, st
tu)
Organ Culture
- Remove the two mesothoracic legs of fifth ins tar nymphs of the cockroach Leucophaea maderae at the trochanter-femoral joint 2 days after molting
- Open the stumps from 10 to 21 days after removal of the legs, and
dissect out the regenerating tissues
Results
According to Marks and Reinecke (1964), in a culture of regenerating
tissues of the legs of L. maderae, active cell migration, pinocytosis, and
phagocytosis of cells in the explants were observed. In addition, the production of vesicles by explanted tissues was seen. These vesicles were
hollow, spherical structures that, when mature, consisted of a single layer
of thin epithelial cells, which became interlaced with attenuated fiberlike cells. Later, Marks and Leopold (1970) obtained the chitinous cuticle
on the explants. However, the cuticle formed in vitro remained thin and
only partially developed. The heavy layer of endocuticle was largely absent. Bulliere and Bulliere (1977) cultured the regenerating appendage
tissues of embryonic legs of Blaberus craniifer in a salt mixture (NaCI
12.65 g, KCI l.61 g, and CaCl 2 l.26 g in 1,000 ml H 2 0) containing 25%
bovine serum. The embryo of this cockroach has two molting cycles,
and regeneration of appendage is possible only during the first molting
cycle. When tarsus segments of the embryonic appendage were removed
and the appendage was cultured, regeneration took place inside the cuticle of the stump. The addition of molting hormone to the medium inhibited regeneration.
Subprotocol 19
Tumor Tissues of DrosophilQ (Diptera)
Materials
Equipment - Dissecting microscope
- Maximov slide
- Forceps
- Needles
- Pasteur pipettes
- Culture vessel
Solutions - Ringer's solution
- Culture medium (Kuroda and Tamura's medium; see Appendix 1)
Animals - Aseptically reared larvae of D. melanogaster (preferably strain v tu, st
tu)