Good cell culture: Jinma tells me! -Huaqiang Electronic Network

Cell culture cell culture technology is also called cell cloning technology, and the regular term in biology is cell culture technology. Cell culture is an indispensable process for bioengineering technology and bioclone technology. Cell culture itself is a large-scale cloning of cells. Cell culture technology can be cultured from one cell to a simple single cell or a very small multi-cell, which is an essential part of cloning technology, and cell culture itself is a clone of cells. A large number of cells or their metabolites are obtained by cell culture. Because biological products are derived from cells, it can be said that cell culture technology is the core and most basic technology in biotechnology. So if you want to cultivate good cells, Shanghai Jinma tells you where you are, let's take a look at our experience!
1. Ensure that all laboratory materials are aseptically cross-contaminated as a major 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. Careful handling of the fragility of your culture's 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 STR profiling on a regular basis to confirm the identity of the cell line.
3. Properly thaw cells before use. 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. The cell culture process using logarithmic growth phase has three stages: 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. Do not allow cells to completely confluent before passage. Confluence 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 In the cell culture process, the medium is the most important factor in controlling product quality, yield and cost. 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. Evaluating the quality of water when formulating 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 the cells to optimize the media. The used media is used to help identify the metabolic rate of the essential components, 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 the passage of adherent cells by digesting the proteins 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, which is very harmful to the experiment.

In short, what the cell needs to provide, the truth is that it takes time to really do this. People have not enough understanding of the cell's life cycle control mechanism. Although cancer cells are also from normal cells, they have not been Knowing exactly why cancer cells are difficult to stop the harmful divisions that have already started, despite the long-term research results, the basic conditions required for in vitro cell culture are the following cellular physiological conditions. My company

It can be passaged for a long time, maintains activity, and is easy to monitor and detect structural functions and life activities.

Culture conditions can be artificially controlled to study the effects of cellular metabolism, physical, chemical, and biological factors. It can be used to study changes (variation, differentiation) of living cells by various observation and detection methods. The expression of subcellular structures and metabolic molecules can be carried out at the cellular level. The scope of research and the source of cells are wide and the selectivity is wide. Different generations of cells can be preserved for a long time, and different conditions and methods of the same generation can be studied, and the dynamic changes of different generations can be observed. It is low in cost, economical, low in cost, and can be cultivated in large quantities, which is conducive to the production of biological products.