Trypsin EDTA Solution contains 0.25% trypsin and 0.02% EDTA. The digestion solution is filtered and sterilized, and can be directly used for the digestion of cultured cells or the dissociation of some tissues. This product has the characteristics of convenience and speed, and most adherent cells can be digested after about 1 minute of room temperature digestion.
Storage
Long term storage at -20 ℃ for 2 years. Once melted, store at 4 ℃ for 6 months to avoid repeated freeze-thaw.
Operation method
1. Digestion of adherent cells:
a) Suck off the culture medium and wash the cells once with sterile PBS to remove residual serum.
b) Add a small amount of pancreatic enzyme cell digestion solution, slightly cover the cells, and let it sit at room temperature for 30 seconds to 2 minutes. The digestion time of different cells varies.
c) Under a microscope, the cells showed significant contraction, and a significant change in cell morphology was observed at the bottom of the culture vessel with the naked eye; Or blow the cells with a gun and find that they can just be blown down. At this point, remove the digestive fluid of pancreatic enzyme cells. Add complete cell culture medium containing serum and blow down the cells, which can be directly used for subsequent experiments.
d) If indigestion is found, add pancreatic enzyme cell digestion solution for re digestion.
e) If it is found that the digestion time of cells is too long and there is no time to blow the cells, some of the cells have directly fallen off the bottom of the culture vessel, and the cells are directly blown down using pancreatic enzyme cell culture medium. Centrifuge 1000-2000g for 1 minute, precipitate cells, remove pancreatic enzyme cell digest as much as possible, and then add complete culture medium containing serum to resuspend cells, which can be used for subsequent experiments.
2. Digestion of tissues:
The time required for digestion varies greatly among different tissues, and it is usually advisable to fully disperse the tissue after digestion.? Precautions? 1. During the use of pancreatic enzyme cell digestion solution, special attention should be paid to avoiding bacterial contamination of the digestion solution. 2. The digestion time of pancreatic enzyme cells should not be too long, otherwise the growth status of cells will be poor after laying the board.