Making cells better adhere to the wall is the key to ensure their normal growth in the later stage. Sometimes we encounter the situation that the cells in the cell culture flask are not easy to adhere to the wall. The following methods can better promote the growth of cell adherence.
1. Increase inoculation density:
Cells mainly grow by attaching to the surface of the support through the extracellular matrix secreted by themselves. If the cell adhesion is not good, it may be that the number of inoculations is too small to secrete enough extracellular matrix. You can increase the inoculation density to improve adhesion Effect.
2. Increase the adhesion factor:
Serum contains adhesion-promoting factors, but in the serum-free culture system, due to the lack of this component, the cell adhesion effect will be greatly reduced. Some serum can be added appropriately to promote cell attachment.
3. Improve the quality of inoculated cells:
The state of the cells themselves will also affect their adherence effect. Make sure that the cells are in good condition before inoculation, and do not use aged cells.
4. Select consumables processed by TC:
Cell culture flasks are commonly used consumables for adherent cell culture. There are two types of consumables: TC-treated and non-TC-treated surfaces. When selecting consumables, choose products that have been treated with TC. Such products introduce hydrophilic Agglomerates are more conducive to cell growth.
In addition, poor cell adhesion may also be caused by the introduction of other sources of pollution, such as bacteria, fungi, mycoplasma, etc. Operators should pay attention to strictly implement aseptic procedures to avoid contaminating cells.
Cell vacuolation refers to the appearance of vacuoles (vesicles) of different sizes in the cytoplasm and nucleus of degenerated cells, and the cells are honeycombed or reticular. There are many reasons for this situation. We can minimize the vacuolation of cells in the cell culture flask through daily operations.
1. Confirm the cell status:
Determine the state of the cells before culturing the cells, and try to select cells with an earlier algebra for culture to avoid vacuoles due to aging of the cells during the culture process.
2. Determine the pH value of the culture medium:
Confirm the suitability of the pH of the culture medium and the pH required by the cells to avoid affecting cell growth due to inappropriate pH.
3. Control trypsin digestion time:
When subculture, choose the appropriate concentration of trypsin and select the appropriate digestion time for digestion, and avoid excessive blowing and blowing to produce too many bubbles during operation.
4. Observe the cell status at any time:
When culturing cells, observe the state of cells in the cell culture flask at any time to ensure that the cells require sufficient nutrients and avoid vacuolation of cells due to nutrient deficiency.
5. Try to use fetal bovine serum with good quality and regular channels, because such serum is rich in nutrients and has few exogenous stimuli, which can effectively avoid such problems.
Through the above operations, the vacuolation of cells in the cell culture flask can be reduced. In addition, the aseptic requirements must be strictly implemented during operation to reduce the possibility of various contaminations. If the cells are found to be contaminated, discard them in time to avoid adverse effects on subsequent experiments. influences.