How to Rescue Cells with Slow Growth and Poor Adhesion in Passages?

Different kinds of cell products are fundamental to our research experiments. A seemingly insignificant cell holds sway over the success or failure of our experiments. What may seem like minor details regarding cells are, in fact, pivotal in experiments. If your cells exhibit slow growth or detachment from the culture vessel, it's crucial to take heed. These signs indicate poor cell health and necessitate immediate action to rescue them, otherwise, the cells may be on the brink of death.

Issue 1: Slow Cell Growth

Slow cell growth is a common issue encountered during cell culture. Typically, cell viability and concentration serve as indicators of cell health. When cells exhibit slow growth, both viability and concentration may decline linearly.

 Main Causes and Solutions for Slow Cell Growth:

1. The cells are not adapting to a newly prepared culture medium or to a change in serum

Solution: The ideal approach is to select the most appropriate culture medium. However, if situations don't allow for this, when switching mediums, it's advisable to incrementally introduce the new medium instead of abruptly replacing the old one. Begin with a quarter, then half, followed by three-quarters, until eventually transitioning to the full amount. Furthermore, conduct multiple passages to facilitate the gradual adaptation of the cells.

2. Insufficient Cell Density

Some cells are density dependent cells, and low density can lead to slow growth. It could also be due to inadequate secretion of growth-promoting factors or poor cell-cell adhesion.

Solution: Increase the seeding density of the cells. Transfer the cells from large culture dishes to smaller ones. Additionally, consider increasing the concentration of serum in the culture medium appropriately.

3. Changes in Culture Environment

The temperature, CO2 concentration, and overall cell culture conditions within the incubator should be optimal for cell growth. Generally, the recommended temperature is 37°C, CO2 concentration should be maintained at 5%, and the pH level should range between 7.2 to 7.4. It's also crucial to consider factors such as osmotic pressure. It's important to ensure these conditions are properly calibrated and maintained, and avoid unnecessary alterations that may disrupt the optimal growth environment for the cells.

4. Contamination, such as mycoplasma, bacteria, fungi, etc

Solution: Cell contamination can arise from various sources. On one hand, it's crucial to adhere to strict aseptic techniques and maintain a clean environment. On the other hand, in the event of contamination, prompt identification and treatment are necessary. Additionally, it's advisable to use high-quality cell lines and culture media as a preventive measure, as this is the best approach to minimize the risk of contamination.

5. Changes in Cell State, such as aging

Solution: There are no reversible methods for addressing changes like cellular aging. The only recourse is to promptly replace the aged cells with fresh ones.

Additionally, in the case of primary cells, they tend to exhibit slower growth rates. If the medium is not replenished in a timely manner, leading to insufficient nutrient supply, cells may also experience slow growth. When culturing cells in flasks with screw caps, it's important to slightly loosen the cap to ensure proper ventilation. It's also essential to maintain clean air within the CO2 incubator.

Indeed, provided that the conditions support cell growth and the cells are at ease and unconfined in their "little homes," they will undoubtedly proliferate rapidly, and their morphology will progressively become more "beautiful."

Issue 2: Cells Not Adhering to the Surface

Usually, in regular conditions, following a period of cell culture, actively growing cells attach to the surface, thriving in their environment, while those that haven't survived or are of different cell types might remain suspended. If your cells aren't adhering to the surface, it's highly probable that they haven't survived.

Causes and Solutions for Cells Not Adhering to the Surface:

1. Over-digestion with trypsin:

Solution: Reduce the digestion time with trypsin or decrease the concentration of trypsin.

2. Mycoplasma contamination:

Solution: Isolate the culture, test for mycoplasma contamination, and clean the incubator and racks. If mycoplasma contamination is detected, discard the culture.

3. Unclean bottom of the culture vessel:

Solution: Switch to new culture dishes or bottles.

4. Incorrect or expired preparation and storage of digestion or culture media:

Solution: Reconfigure the digestion or culture media.

5. Cellular senescence:

Solution: Similarly, there are no reversible methods. Replace the cells with fresh ones promptly.

6. Initial seeding density too low or too high:

Solution: Seed an appropriate number of cells based on their characteristics.

In summary, cell culture requires patience and gentleness; otherwise, cells may not behave as expected. As mentioned earlier, using high-quality cell lines and culture media is the best way to minimize contamination and ensure optimal cell growth.

We recommend utilizing Pricella's high-quality culture media, and auxiliary reagents.

We provide a wide range of cell culture media and reagents, offering nearly a thousand varieties. Our rigorous quality control system guarantees adherence to quality standards and stability. Additionally, if you encounter any quality concerns with our products, we offer unconditional after-sales service, ensuring a stress-free experience for you every step of the way.

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