Tips for Suspension Culture of Adherent Cells

How to suspend adherent cells for culture? Poloxamer should have its perfect shear force as its primary advantage. Why do we suspend adherent cells for culture? There are several major benefits:

1. Reduce excessive use of consumables

2. Able to obtain the maximum number of cells

3. Maintain consistency of extracted proteins

2、 How can adherents be domesticated into suspended cells?

For adherent cells (2-4 weeks required):

1. When the cell reaches its maximum density, adaptive training begins;

2. Pancreatin digestion of adherent cells, 350 × Centrifuge for 5 minutes to collect cells, and resuspend with serum-free medium containing 5% serum to achieve a cell density of 4 × 10 ^ 5 cells/ml.

3. Cultivate cells to maximum density;

4. Repeat steps 2-3 and sequentially add 2%, 1%, 0.5%, and 0.1% serum to serum-free medium for cell culture; If the cell survival rate is below 80%, it is necessary to restore to the previous serum level or adopt a more refined decreasing content.

5. Until cells are completely cultured using serum-free medium.

Qida Biology provides a suspension ™ A series of culture media, formulated with serum free five animal components, without animal protein, composed of small and large molecule compounds and a small amount of hormones, suitable for large-scale reactor culture. Serum free culture has the following advantages:

The consistency of serum free culture medium batches is high, which can eliminate the need for pre screening the culture medium.

The cost of serum free culture medium has decreased, and the price fluctuation is not significant. If serum is used, it is easy to be affected by the surrounding environment (such as a decrease in serum production during dry weather), leading to adjustments in serum prices.

Serum free culture medium does not contain mitogen inhibitors and can promote cell proliferation.

Serum free culture medium can reduce the possibility of microbial contamination by viruses, fungi, mycoplasma, and other microorganisms.

The components of serum free culture medium are stable and can be prepared in large quantities.

Cultivating cells in serum free medium can simplify the process of purification and identification of various cell products.

Suspension ™ CHO (XF) cell suspension culture medium

The cell culture method from serum-containing to serum-free is as follows:

For suspended cells (2-4 weeks required):

1. Start adaptive training when the cell density reaches its maximum;

2. Add serum-free medium to reduce the ratio of serum-free medium to normal medium to 1/2 of the original one;

3.When the cell density reaches its maximum, add serum-free culture medium to reduce the cell density to 1/5 of the original value;

4. Cultivate cells. If the cell survival rate is greater than 85% when the maximum density is reached, and the time to reach the maximum density is similar to before, then the cells can continue to undergo adaptive training; If the cell shows slow growth or reduced survival rate, repeat steps 2-3 three times;

5. The proportion of serum-free medium can be gradually increased until cells can fully grow in serum-free medium.