108
Methods for Setting Up Primary Cultures Specific to Animal Groups
• • Results
Fig. 1. Spherical vesicles formed in the
primary culture of embryonic tissues of
the German cockroach Blattella
germanica. x 200 From Kurtti, T.J. and
Brooks, M.A., "Isolation of cell lines from
embryos of the cockroach, Blattella
germanica", In Vitro, Vo1.13, pp. 14, Copyright © 1977 by the Society for In Vitro
Biology (formerly the Tissue Culture Association). Reproduced with permission
of the copyright owner. (1977)
The primary culture contains single cells, clusters of cells, and fragments
of embryos. The overall growth of the primary culture is slow. The migrated cells may form hollow spherical vesicles that continue to grow
(Fig. O. It will take many weeks before confluency is established. When
cell numbers have increased sufficiently, a subculture can be made; however, during the early stages of subculturing, cell growth will be slow.
The hollow spherical vesicles can be subcultured also. Continuously growing cell lines may be obtained from such cultures by repeating the subculturing process. Several continuous cell lines have been obtained from
the American cockroach Periplaneta americana (Landureau, 1968) and
the German cockroach Blattella germanica (Kurtti and Brooks, 1977).
I
Subprotocol 2
Hemocytes
• • Materials
Equipment - Beakers
- Culture vessels
- Scissors
- Pasteur pipettes
Solutions - Culture medium (S-20; see Appendix 1)
Chemicals - Detergent (neutral detergent for kitchen use)
- Sodium hypochlorite
Animals - Larval or adult cockroaches
• • Procedure
Sterilization l. Wash the cockroaches in a solution of 20% detergent.
2. Surface sterilize the cockroaches by dipping them into a solution of
sodium hypochlorite.
3. Wash the cockroaches with sterile distilled water.
Methods for Setting Up Primary Cultures Specific to Animal Groups
• • Results
Fig. 1. Spherical vesicles formed in the
primary culture of embryonic tissues of
the German cockroach Blattella
germanica. x 200 From Kurtti, T.J. and
Brooks, M.A., "Isolation of cell lines from
embryos of the cockroach, Blattella
germanica", In Vitro, Vo1.13, pp. 14, Copyright © 1977 by the Society for In Vitro
Biology (formerly the Tissue Culture Association). Reproduced with permission
of the copyright owner. (1977)
The primary culture contains single cells, clusters of cells, and fragments
of embryos. The overall growth of the primary culture is slow. The migrated cells may form hollow spherical vesicles that continue to grow
(Fig. O. It will take many weeks before confluency is established. When
cell numbers have increased sufficiently, a subculture can be made; however, during the early stages of subculturing, cell growth will be slow.
The hollow spherical vesicles can be subcultured also. Continuously growing cell lines may be obtained from such cultures by repeating the subculturing process. Several continuous cell lines have been obtained from
the American cockroach Periplaneta americana (Landureau, 1968) and
the German cockroach Blattella germanica (Kurtti and Brooks, 1977).
I
Subprotocol 2
Hemocytes
• • Materials
Equipment - Beakers
- Culture vessels
- Scissors
- Pasteur pipettes
Solutions - Culture medium (S-20; see Appendix 1)
Chemicals - Detergent (neutral detergent for kitchen use)
- Sodium hypochlorite
Animals - Larval or adult cockroaches
• • Procedure
Sterilization l. Wash the cockroaches in a solution of 20% detergent.
2. Surface sterilize the cockroaches by dipping them into a solution of
sodium hypochlorite.
3. Wash the cockroaches with sterile distilled water.