Suspension™ MDCK Medium(XF)

Suspension™ MDCK series suspension medium is a medium that supports BHK cell suspension culture. It is suitable for MDCK suspension culture and large-scale reactor culture. It consists of more than 100 compounds and does not contain protein, but contains a small amount of hormones. It is used Ferric citrate and other alternatives to the application of transferrin in cell growth, using the Suspension™ series of suspension cell culture media, the maximum cell density that can be cultured is above 10×10^6 cells/ml. The culture medium can be used to transfect cells to improve For transfection efficiency, antibiotics are in independent small packages and do not need to be used to improve transfection efficiency during cell transfection. 

Medium composition: 500ml Suspension™ MDCK (XF) suspension cell culture medium (Qida Biotech: SP01), 5ml G /A (Qida Biotech: SD0038) 

The advantages of serum-free culture medium are as follows: 

● Serum-free culture medium has high batch consistency, which eliminates the need for pre-screening of the culture medium.
● The cost of serum-free medium is reduced and the price fluctuates little.If serum is used, it is easily affected by the surrounding environment (for example, when the weather is dry, serum production decreases), causing the serum price to be adjusted.
● Serum-free medium does not contain mitogen inhibitors and can promote cell proliferation.
● Serum-free medium can reduce the possibility of microbial contamination by viruses, fungi, mycoplasma and other microorganisms.
● The components of serum-free medium are stable and can be prepared in large quantities.
● Cultivation of cells in serum-free medium can simplify the procedures for purification and identification of various cell products.

Operation steps: 

The adaptive training method of cells from regular culture medium to serum-free medium is divided into 2 types according to the type of cells: 

For suspension cells (it takes 2-6 weeks):
1. Start adaptive training when the cell density reaches the maximum ;
2. Add serum-free medium to make the ratio of serum-free medium and normal medium to 1/2 of the original.
3. When the cell density reaches the maximum, add serum-free medium to make the cell density to 1/2 of the original. 5;
4. Culture cells. If the cell survival rate is greater than 85% when the cells reach the maximum density, and the time to reach the maximum density is similar to before, then the cells can continue adaptive training; if the cells show slow growth or reduced survival rate , then steps 2-3 need to be repeated 3 times;
5. The proportion of serum-free medium can be gradually increased until the cells can completely grow in serum-free medium.

For adherent cells (it takes 2-6 weeks):
1. When the cells reach the maximum density, start adaptive training;
2. Digest the adherent cells with trypsin, centrifuge at 350×g for 5 minutes to collect the cells, and use serum-free solution containing 5% serum. Resuspend the culture medium to make the cell density reach 4×10^5 cells/ml.
3. Culture the cells until they reach the maximum density;
4. Repeat steps 2 - 3 and add 2%, 1%, 0.5% and 0.1% serum to the serum-free medium in sequence for cell culture; if the survival rate of the cells is lower than 80%, then it is necessary to return to the content of the previous serum, or adopt a more detailed decreasing content.
5. Culture cells in serum-free medium until completely.

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