54
Methods for Setting Up Primary Cultures Specific to Animal Groups
Subprotocol 5
Fat Bodies
Fig. 4. Cultured midgut cells of
Heliothis virescens. C, differentiated
columnar cells; D, differentiating
cells; G, differentiated goblet cells; S,
stem cells; Sc, secretory cells. x800.
(Figure courtesy of Dr. Marcia Loeb,
United Stats Department of Agriculture, Insect Biocontrol Laboratory,
Beltsville, MD, USA)
Fat bodies are known as organs in which carbohydrates are stored as an
energy source, and in which protein synthesis takes place. Therefore, if a
continuous cell line is obtained from fat body tissues, it will be useful in
the study of the biochemistry of metabolism .
• • Materials
Equipment The equipment to be used is the same as that given for subprotocol 1.
Solutions - Physiological solution
- Culture media
Animals - Matured final instar larvae of lepidopteran insects
Sterilization
and cflSSeCtion
Culture
set-up
• • Procedure
Follow the instructions given for sterilization, and steps 1-4 for dissection as listed in subprotocol 1.
1. Take out as many major lobes of the fat bodies as possible.
2. Remove as many attached trachea as possible.
3. Cut the fat bodies into pieces smaller than 1 mm.
4. Wash the fragmented fat bodies with physiological solution and transfer to culture medium.
5. Transfer the fragments, with the culture medium, into culture vessels.
6. Culture at 25°C.
Methods for Setting Up Primary Cultures Specific to Animal Groups
Subprotocol 5
Fat Bodies
Fig. 4. Cultured midgut cells of
Heliothis virescens. C, differentiated
columnar cells; D, differentiating
cells; G, differentiated goblet cells; S,
stem cells; Sc, secretory cells. x800.
(Figure courtesy of Dr. Marcia Loeb,
United Stats Department of Agriculture, Insect Biocontrol Laboratory,
Beltsville, MD, USA)
Fat bodies are known as organs in which carbohydrates are stored as an
energy source, and in which protein synthesis takes place. Therefore, if a
continuous cell line is obtained from fat body tissues, it will be useful in
the study of the biochemistry of metabolism .
• • Materials
Equipment The equipment to be used is the same as that given for subprotocol 1.
Solutions - Physiological solution
- Culture media
Animals - Matured final instar larvae of lepidopteran insects
Sterilization
and cflSSeCtion
Culture
set-up
• • Procedure
Follow the instructions given for sterilization, and steps 1-4 for dissection as listed in subprotocol 1.
1. Take out as many major lobes of the fat bodies as possible.
2. Remove as many attached trachea as possible.
3. Cut the fat bodies into pieces smaller than 1 mm.
4. Wash the fragmented fat bodies with physiological solution and transfer to culture medium.
5. Transfer the fragments, with the culture medium, into culture vessels.
6. Culture at 25°C.