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Methods for Setting Up Primary Cultures Specific to Animal Groups
Culture 1. Cut the embryos into several pieces.
Fig. 1. Cell migration from explanted embryonic tissue of the
May-beetle Anomala costata. EP, a
cell sheet of epithelial cell-like
cells; EX, explant; H, hemocytelike cells. x 150
set-up 2. Transfer the embryo fragments into a culture vessel with the culture
medium .
• • Results
Cells will migrate from the cut opening of the embryo fragments (Fig. 1).
The migrated cells may proliferate if the culture conditions are favorable. If cells proliferate quickly, a continuous cell line may be obtained by
repeating subculture of the cells. The cell line from embryos may differentiate to a certain cell type during repeated subculturing (Mitsuhashi,
1989). Several continuous cell lines have been established from the embryonic tissues of beetles (Crawford, 1982; Lynn and Stoppleworth, 1984;
Mitsuhashi, 1989; Lynn, 1995).
Subprotocol 2
Hemocytes
• • Materials
Equipment - Centrifuge and centrifuge tube
- Needles
- Scissors
- Pasteur pipette
Solutions - Physiological solution
- Culture medium
Animals - Larvae of coleopteran insects