step:
1. Prepare the required number of Feeder cell plates the previous afternoon;
2. Remove the medium from the 4 °C refrigerator about half an hour in advance and put it at room temperature (or in a 37 ° C incubator);
3. Take the ES cell culture dish and the Feeder cell culture dish out of the incubator (conventional observation under phase contrast microscope, Feeder cells should grow well, the cells cover about 95%, at least 90% of the culture dish area); ES cells should It grows well and is close to the Petri dish. The cell colonies are uniform in size and uniform in density. The colonies are regular in shape, oval in shape, smooth in edge, smooth in surface, dense in cell growth, large in nuclear size and low in cytoplasm. Shanghai Chuangsai Technology Co., Ltd. provides 6.0CM. Disposable cell culture dish, flat-bottom sterilization, suitable for cell adherent growth, commodity number: C81-TCD010060-600 pcs/carton, price 652.5 yuan.
4. Aspirate the liquid in the ES cell culture dish, discard it, wash it with PBS 1 mL, add trypsin solution for about 30 seconds, carefully aspirate the trypsin solution, discard it; then apply for 1-5 minutes, from time to time Observe that when the cell layer (a layer of white dirty things on the bottom of the dish) cracks and begins to slide down, add the medium and moderately blow it (depending on the degree of digestion and the thickness of the tube, it is not obvious). Until the cell clumps; average into the Feeder culture dish (drop, so as not to blow the Feeder cells off), add additional medium to about 5 ml (drop, so as not to blow the Feeder cells off; if 3.5cm culture dish, add to about 3ml), make a label; Shanghai Chuangsai Technology provides E30-LW300LT biological microscope, commodity number: E30-LW300LT biological microscope, inquiry.
5. Shake the culture dish horizontally before, after, and so on, so that the ES cells are evenly distributed in the culture dish and placed in the incubator to continue the culture.
Excellent performance of CANSPEC, cytokines, fetal bovine serum, electrophoresis equipment, standard products, chemical and chemical, scientific instruments, laboratory supplies, Shanghai Chuangsai, mass products special promotions, welcome to inquire!
Towel Kitchen Towels,Absorbent Kitchen Towel,Towels Hand Towel,Hand Towel
Baoda Paper Enterprise Co., Ltd. , https://www.baodatissue.com