Bombyx mori Cell Culture The culture using cells obtained directly from the body is called primary culture. This culturing process mainly adopts the method of aseptic operation to take out the tissues (organs) from the animal body, undergo enzyme digestion treatment to disperse them into single cells, and then cultivate them under artificial conditions to continuously grow and reproduce. Primary culture is the first step in establishing various cell lines. Since the primary cultured cells have just been separated from the living tissue, they can reflect the living state of the organism to a certain extent.
1. The purpose of the experiment
1. Understand the basic method and operation process of primary cell culture.
2. Familiar with the observation method of primary cultured cells.
Second, the experimental principle The cultivation of cells obtained directly from the body is called primary culture. This culturing process mainly adopts the method of aseptic operation to take out the tissues (organs) from the animal body, undergo enzyme digestion treatment to disperse them into single cells, and then cultivate them under artificial conditions to continuously grow and reproduce. Primary culture is the first step in establishing various cell lines. Since the primary cultured cells have just been separated from the living tissue, they can reflect the living state of the organism to a certain extent. The primary culture technology can be used to study the proliferation, inheritance, mutation, differentiation and dedifferentiation, malignant transformation and demalignant transformation of various types of cells in vitro. Primary culture can be divided into tissue block culture method and digestion method.
3. Experimental instruments, materials and reagent instruments and utensils: biochemical incubator, inverted microscope, ultra-clean workbench, pressure cooker, constant temperature water bath, centrifuge, ophthalmic scissors, ophthalmic forceps, wax plate, centrifuge tube, culture bottle, gauze Petri dishes, micropipettes, pipettes, pipettes, alcohol lamps, alcohol cotton balls, test tube racks, etc.
Material: Silkworm eggs.
Reagents: D-Hanks solution (that is, Hans solution without calcium and magnesium ions), 0.25% trypsin solution, TC-100 medium (GIBCO company), 2% Xinjieer disinfection solution, Grace medium (GIBCO company), fetus Bovine serum.
4. Experimental steps
1. Sterilize silkworm eggs with 2% Xinjieer solution, then disinfect with 75% alcohol cotton, and dry;
2. Disinfect the double-sided blade and ophthalmic scissors with alcohol cotton;
3. Take an appropriate amount of culture medium in a larger Petri dish, use a double-sided blade to cut the egg shell, try to maintain the integrity of the embryo, use a short pipette to suck the medium to blow out the embryo in the egg;
4. Take the appropriate amount of culture medium in a small petri dish, and use a short pipette to suck the blown out complete embryos into the small petri dish.
5. Repeat 4 steps twice;
6. Finally, suck the embryo into a small beaker containing a small amount of culture medium, and use an ophthalmic scissors to cut the embryo to a suitable size;
7. Treat with 0.25% trypsin for 10-20 minutes to form a flocculent precipitate, remove the trypsin with a long pipette, rinse the precipitate with fresh medium 1-2 times, add an appropriate amount of medium to a small beaker, and use a long pipette to break up the precipitate The suspension is made, and the medium and fetal bovine serum are added at a ratio of 5: 1;
8. Pipette the suspension into a 25ml cell bottle and place it in a 28 ° C biochemical incubator. Change the medium on the third day, and treat it according to the cell growth.
Minimalist design, smooth frame processing, a good companion to accompany the baby, the product design is lightweight, convenient for the baby to carry, and draw whenever and wherever they want. Say goodbye to ink dust and play with the 3D special effect Drawing Board. We create every detail with our hearts and feel every ingenuity behind it.
RGB Drawing Board,fluorescent drawing board,magic fluorescent drawing board
Guangdong Jishengke Industrial Co.,Ltd , https://www.jskpad.com