Analysis of Reasons for Cell Detachment in Cell Factories

Effective cell adhesion is a critical step in the successful cultivation and expansion of cells in cell factories. However, sometimes we face issues where cells fail to adhere, which can be caused by various factors. Here are some common reasons and their solutions:

Effective cell adhesion is a critical step in the successful cultivation and expansion of cells in cell factories. However, sometimes we face issues where cells fail to adhere, which can be caused by various factors. Here are some common reasons and their solutions:

1. Excessive Trypsinization
Trypsin is used for cell dissociation and passage, but if the treatment time is too long or the concentration is too high, it may damage cell membrane proteins, affecting the cells' adhesion ability. To avoid this, trypsinization time can be shortened or the concentration reduced, minimizing cell membrane damage and thereby improving cell adhesion rates.

2. Microbial Contamination
Microbial contamination, especially mycoplasma infection, is another common issue. The presence of microorganisms can severely interfere with cell function and growth, impacting their adhesion ability. Once microbial contamination is detected, immediate isolation and mycoplasma testing should be conducted. If mycoplasma infection is found in the cell factory, contaminated cultures must be thoroughly removed to prevent further spread.

3. Alkaline Culture Medium pH
An excessively high pH in the culture medium may create an unfavorable environment for cells, affecting their normal adhesion. In such cases, the pH can be adjusted by adding sterile acetic acid solution or injecting sterile CO2 to maintain an appropriate acid-base balance, thereby promoting cell adhesion and growth.

4. Cell Aging
As cells undergo multiple passages, they may gradually age, losing their adhesion and normal growth characteristics. To prolong the effective lifespan of cells, it is recommended to regularly update the cell seed stock and add an appropriate amount of serum during passages to help cells recover and rebuild their adhesion properties.

5. Improper Initial Cell Seeding Density
The initial seeding density of cells is crucial for the formation and quality of the extracellular matrix. A density that is too low may fail to form an effective extracellular matrix, affecting cell adhesion. To address this, the composition of the culture medium can be optimized, using media rich in factors that promote cell adhesion, ensuring stable cell attachment and growth.

The successful operation of cell culture experiments relies on the health and effective adhesion of cells during the cultivation process. By identifying and addressing common issues such as excessive trypsinization, microbial contamination, alkaline culture medium pH, cell aging, and improper cell seeding density, the stable operation and high-efficiency production of cells in cell factories can be ensured, providing a solid foundation for the development of biopharmaceuticals, vaccine preparation, and other biotechnological fields.