Serum-free cell freezing medium instructions

1. Product Overview

Serum-free cell freezing solution can be used for mammalian cells, especially for the cryopreservation of various immune cells such as PBMC and NK. The cell viability after cryopreservation and resuscitation is over 90%. It is serum-free, free of xenobiotic components, and has limited chemical composition. It does not require complex programmed cooling. It is ready-to-use, does not require preparation, and is easy to use.

2. Packing Specifications

Item No.: FCS-2401; Specification: 100mL/bottle

3. Storage conditions and shelf-life

Storage conditions: 2-8℃; validity period: 12 months

4. How to operate

Cryopreservation of cells:

1. Observe the cell status, count the cells, and determine the cell viability (greater than 90%) before freezing.

2. Collect the cells to be frozen in a centrifuge tube, centrifuge at 1000~ 1500rpm for 5 min, and discard the supernatant.

3. Adjust the cell freezing density as needed and slowly add an appropriate volume of serum-free cell freezing solution stored at 4°C to resuspend the cells. Note: Choose the appropriate freezing density according to different cells and experimental arrangements. The recommended freezing density is 5.0 × 106 ~3.0 × 107 cells/mL.

4. Aliquot the cell suspension into cryopreservation tubes, tighten the tube caps, and label them.

5. Place the cryovial in a programmed cooling box (precooled at 4°C) , then place it in a -80°C refrigerator overnight, and transfer it to a liquid nitrogen tank the next day .

Cell Recovery:

1. Preheat a water bath to 37°C in advance.

2. Take out the cell cryopreservation tube from the liquid nitrogen tank, quickly place it in a 37°C water bath and shake it gently. Take it out when the ice crystals in the cell suspension are about to completely disappear as observed by naked eyes.

3. Clean the tube with 75% alcohol, quickly transfer the cell suspension into the culture medium (no preheating required) and mix thoroughly (if the frozen cell suspension is 1 mL, add it to 9 mL culture medium).

4. 1000rpm~1500rpm, centrifuge immediately for 5 min, discard the supernatant, and add fresh culture medium preheated at room temperature.

5. As needed, cells of appropriate density are seeded into a suitable culture container and transferred to an incubator for culture.

5. Precautions

1. After adding this product to cells, the storage time at room temperature should be minimized and the cells should be moved to -80℃ for storage as soon as possible.

2. Cryopreservation of cells in the logarithmic growth phase can help improve the survival rate of revived cells.

3. This product contains DMSO. For some cells that are sensitive to DMSO, it is recommended to conduct a preliminary experiment first.

5. This product contains DMSO. For some cells that are sensitive to DMSO, it is recommended to conduct a preliminary experiment first.