Cells growing in monolayers proliferate, fuse into sheets, and grow all over the surface of the cell vial. Some cells can be maintained in the plateau phase for days to weeks by frequent fluid changes; while many other cell lines require trypsin treatment, passaging culture to survive, and treatment with trypsin to separate cells from the surface of the growth to facilitate passaging culture. This experiment was derived from Cell Lab Guide (Previous) by Peitang Huang.
Operation method
Inoculation of monolayer cells in cell vials
Principle
Cells growing in monolayers proliferate, fuse into sheets, and grow all over the surface of the cell vial. Some cells can be maintained at a plateau for days to weeks with frequent fluid changes, while many other cell lines require trypsin treatment and passaging to survive, and trypsin treatment detaches the cells from the surface of the growth material to facilitate passaging. The latter cell lines are less likely to show contact inhibition and continue to proliferate, and when they reach their limit, the cells are detached from the surface of the vials, making it difficult to re-inoculate them diffusely. Move 1) The following steps describe the process of trypsinization of cells (see "Trypsinization of Isolated Cells" below). For more product details, please visit Aladdin Scientific website.
2) Resuspend the cells in culture medium.
It is good practice to count the cells at this point. If the number of passages is to be recorded to identify the growth experience of the cells, the cells can be diluted at a ratio that facilitates the passaging program.
3) Dispense the cell suspension into a cell culture flask or dish containing culture medium of appropriate pH.
The passaged cells are not able to regulate their own PH for about a few hours before they attach to the wall and metabolize again. the passaging period is particularly important.
The pH of the culture medium and the CO2 level in the gas phase of the cell culture flask or dish should be equilibrated before inoculating the cells. Add the culture solution to the culture flask before cell addition, loosely close the lid, and let it stand in the incubator for 10-15 minutes.
4) Mix the cells in the cell culture flask or dish receiving the cells, spread the cell suspension evenly on the surface of the container, cell walling is usually 8~10 hours in the bed, human diploid fibroblasts are often passed on once a week at a ratio of 1:4. The cell multiplication time can be estimated by the following methods: that is, the time of spreading the culture flask again after the full version of cells is passed in 1:2 ratio, or the time of spreading the cells again after the cell multiplication time is passed in 1:4 ratio for two times.