50
Methods for Setting Up Primary Cultures Specific to Animal Groups
Oocytes neither migrate nor multiply. Some migrated cells may multiply by mitosis, and colonies of multiplied cells may be formed. Migrated
cells and multiplied cells are not uniform. The multiplication rate is usually very low at first. The rate increases with advancing culture, because
multiplying cells become predominant. When most of the culture area is
covered by multiplying cells, a subculture is made. Usually, it will take
many months, or even more than 1 year, until a primary culture is able to
be subcultured. Many continuously growing cell lines have been established from pupal ovaries (e.g., Grace, 1962; Mitsuhashi, 1973, 1977).
Subprotocol 2
Testes
Testes are also situated on the dorsal side in the 5th abdominal segment
of larvae. In some species, the testes of both sides unite to form a large
ball that is situated just under the dorsal vessel.
• • Materials
The materials to be used are the same as those given for subprotocol 1.
• • Procedure
The procedure is the same as given in subprotocoll.
• • Results
Cells emerge from the epithelia of testis fragments (Fig. 3). Undeveloped
spermatocysts may grow to some extent; for example, spherical
spermatocysts may enlarge and elongate to become piriform. Isolated
spermatocytes may develop to a lesser extent, and are rather short -lived.
Some cells that have migrated from the epithelia will divide, and these
Fig. 3. Cell migration from the testis
epithelium of the rice stem borer Chilo
suppressalis. x 50
Methods for Setting Up Primary Cultures Specific to Animal Groups
Oocytes neither migrate nor multiply. Some migrated cells may multiply by mitosis, and colonies of multiplied cells may be formed. Migrated
cells and multiplied cells are not uniform. The multiplication rate is usually very low at first. The rate increases with advancing culture, because
multiplying cells become predominant. When most of the culture area is
covered by multiplying cells, a subculture is made. Usually, it will take
many months, or even more than 1 year, until a primary culture is able to
be subcultured. Many continuously growing cell lines have been established from pupal ovaries (e.g., Grace, 1962; Mitsuhashi, 1973, 1977).
Subprotocol 2
Testes
Testes are also situated on the dorsal side in the 5th abdominal segment
of larvae. In some species, the testes of both sides unite to form a large
ball that is situated just under the dorsal vessel.
• • Materials
The materials to be used are the same as those given for subprotocol 1.
• • Procedure
The procedure is the same as given in subprotocoll.
• • Results
Cells emerge from the epithelia of testis fragments (Fig. 3). Undeveloped
spermatocysts may grow to some extent; for example, spherical
spermatocysts may enlarge and elongate to become piriform. Isolated
spermatocytes may develop to a lesser extent, and are rather short -lived.
Some cells that have migrated from the epithelia will divide, and these
Fig. 3. Cell migration from the testis
epithelium of the rice stem borer Chilo
suppressalis. x 50