Cell shaping involves the steps of cell injection, shaping, and obtaining. The cells in the cell culture flask must grow and develop to a certain extent. At this moment, it is necessary to rely on trypsin to digest the cells on the outer wall of the flask and then feel it. In the case of enzymes, the digestion time must be controlled.
T75 cell culture flask
Trypsin belongs to a different class of trypsin, which can degrade proteins on cell membranes fused to cell culture flasks. By dissolving the protein on the corresponding part, the protein at the junction between the proteins is dissolved, and the cell becomes spherical due to the support force of the cell frame of its own internal structure, thereby separating the cells. The level of trypsin digestion is a key factor in cell shaping: excessive digestion increases cell debris, black debris increases, cells will fall off as a whole, seriously jeopardizing cell viability, and some cells float, with the discarded trypsin Outflow; if the digestion is not enough, it is difficult for the cells to be blown down from the inner wall of the bottle, and the continuous blowing and beating will also damage the cell activity.
cell culture flask
Serum protein culture solution bottle
In different institutions or cells, the effect of pancreatic enzymes is not the same. The activity of the pancreatic enzyme dispersed cells is also related to its concentration value, temperature and effect duration. When the pH is 8.0 and the temperature is 37, the efficacy level of the pancreatic enzyme aqueous solution is relatively strong. When applying trypsin, the concentration, temperature and duration should be well controlled to prevent cell damage caused by excessive digestion. Because Ca2, Mg2 and serum protein, protein grams reduce the activity of pancreatin, so when preparing aqueous solution of pancreatin, use BSS without Ca2, Mg2, such as: D-Hanks solution. When stopping digestion, trypsin can be stopped with serum protein cell culture medium or trypsin retarder to stop the effect of trypsin on cells.
culture flask
T25 cell culture flask
In addition, if it is a newly purchased cell, you can use a lower concentration of trypsin to explore the digestion time first, and observe whether the cells become round every minute, and make a good record. All in all, when obtaining the cells in the cell culture flask, it is necessary to control the trypsinization time, so as to effectively maintain the viability of the cells.
