62
Methods for Setting Up Primary Cultures Specific to Animal Groups
Sterilization
and
dissection
Culture
set-up
Fig. 13. Hemocytopoietic tissue of the
swallowtail butterfly Papilio xuthus. x
40
Fig. 14. A wing disc with attached hemocytopoietic
organ (arrow)
(Fig. 13), and the HPOs are present only during the larval stage. It is not
certain whether HPOs produce only prohemocytes or all types of differentiated hemocytes.
•
Materials
The materials used are the same as those given for subprotocol 5 .
• • Procedure
1. Follow the instructions for sterilization given in subprotocol 1, and
step 1-2 for dissection given in subprotocol6.
2. Remove tissues other than the wing discs.
3. The HPOs are a ring-shaped monolayer of cells. They attached to the
inside surface of the wing discs. They are transparent and barely visible when attached to the wing discs. By touching the surface of the
wing discs with a needle, you may recognize the HPOs (Fig. 14).
4. Separate the HPOs from the wing discs with needles.
1. Transfer the HPOs into culture medium.
2. Cut the tissue into at least two.
3. Transfer the fragments, with the culture medium, into culture vessels.
4. Culture at 25°C.
Methods for Setting Up Primary Cultures Specific to Animal Groups
Sterilization
and
dissection
Culture
set-up
Fig. 13. Hemocytopoietic tissue of the
swallowtail butterfly Papilio xuthus. x
40
Fig. 14. A wing disc with attached hemocytopoietic
organ (arrow)
(Fig. 13), and the HPOs are present only during the larval stage. It is not
certain whether HPOs produce only prohemocytes or all types of differentiated hemocytes.
•
Materials
The materials used are the same as those given for subprotocol 5 .
• • Procedure
1. Follow the instructions for sterilization given in subprotocol 1, and
step 1-2 for dissection given in subprotocol6.
2. Remove tissues other than the wing discs.
3. The HPOs are a ring-shaped monolayer of cells. They attached to the
inside surface of the wing discs. They are transparent and barely visible when attached to the wing discs. By touching the surface of the
wing discs with a needle, you may recognize the HPOs (Fig. 14).
4. Separate the HPOs from the wing discs with needles.
1. Transfer the HPOs into culture medium.
2. Cut the tissue into at least two.
3. Transfer the fragments, with the culture medium, into culture vessels.
4. Culture at 25°C.