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They all regard the cell as their own baby.
How is the baby living, and the mood is good,
Being unhappy is something that you should pay attention to every day.
Because I’m worried about being indifferent,
Cell baby is not good,
Even if it is contaminated, the cells are over!
How to make the cells happy, let's look down together!
1. Make sure all laboratory materials are sterile <br>Cross contamination is the enemy of cell culture. Even the slightest pollution can ruin the results of several weeks. Because the incubator is warm and humid, the fungus grows easily, so care must be taken to clean it regularly. In addition, before placing the culture bottles, pipettes, and other related items in the clean bench, wipe them with alcohol to avoid contamination.
2. Carefully handle your culture <br>The fragility of cell culture cannot be overemphasized. Severe shaking, or continuous temperature fluctuations, can have an adverse effect on growth. Make sure the incubator is level, uniform, and away from the electrical instrument. In addition, try to avoid processing multiple cell lines at once, as it may affect the genotype and phenotype of the cells. You should also complete the STR map analysis on a regular basis to confirm the identity of the cell line.
3. Thaw the cells properly before use <br> Although thawing seems to be a simple step, it must be done properly to avoid damaging the cells. Prolonged exposure to high temperatures can cause cells to fail to plate. Therefore, the cryotube should be placed in a 37 ° C water bath for 2 minutes and then diluted with conditioned medium to avoid direct damage from DMSO.
4. Cells in logarithmic growth phase <br>The cell culture process has three phases: stagnation, log phase and plateau, which represent low cell growth, high cell growth and cell-free growth, respectively. The most viable cells are healthy, rapidly dividing, and present in the log phase at 70-80% confluence. 
5. Don't let the cells completely merge before passing
Confluency refers to the proportion of adherent cells occupying the surface area of ​​the culture flask. Complete confluence means that 100% of the surface is covered by adherent cells. This state must be avoided because it means that cells cannot continue to grow. However, it is imperative to use actively growing cells.
6. Choosing the best medium development strategy
Medium is the most important factor in controlling product quality, yield, and cost during cell culture. Media must be tailored to each culture to optimize experimental results. There are several options when deciding which way to develop your media. You can buy off-the-shelf, develop it yourself, or work with another company to develop a specific medium. In this process, you need to consider factors such as time and cost.
7. Evaluate water quality when preparing dry powder media
Liquid media tends to result in higher quality than dry powder media. This may be related to the quality of the water. Because bacteria grow rapidly in water, endotoxin and other contaminants need to be monitored. Commercial companies have the resources to monitor and control bacterial growth, such as filters. Therefore, the use of commercially produced liquid media will be more reliable.
8. Use the metabolic properties of cells to optimize the medium
Analysis of the used medium helps to identify the metabolic rate of the essential ingredients, including amino acids and vitamins. Dynamic metabolic profiles collected from the cell growth phase and the protein production phase can be used to balance the basal medium and add media, directing the focus of the medium and contributing to protein production.
9. Avoid excessive digestion of cells
Trypsin achieves passage of adherent cells by digesting the protein responsible for binding the cells to the container. If the cells are exposed to trypsin for too long, trypsin will begin to cleave cell surface proteins, which affects the ability of the cells to function properly.
10. Do not use antibiotics all the time in the medium.
As cell cultures are more frequently exposed to antibiotics, strains that are naturally resistant to antibiotics will begin to appear. This phenomenon can mask potential pollution.
How do you know if your cells are happy? ——Cell culture precautions
Research students raise cells,