Serum-free rapid cell cryopreservation operating instructions

Serum-free rapid cell cryopreservation solution is commonly used in various animal cell lines. The special formula has an effective increase in cell freezing survival rate and resuscitation vitality. Contains no animal-derived protein, which can reduce the pollution of various viruses, molds and mycoplasma, and ensure the safety of frozen cells. It is suitable for cryopreservation of general cultured cells as well as cryopreservation of serum-free cultured cells and protein-expressing cells.

feature of product
1. High safety, completely using pharmaceutical grade raw materials for production, free of animal components, low possibility of pollution such as viruses, molds and mycoplasma, and higher product quality consistency among batches of products.
2. Cell survival rate is high, no batch difference.
3. Completely frozen solution formula, can be used directly, convenient and simple, can be directly stored in -80 ° C freezer, without the need for time-consuming procedures to cool down (saving time, labor, money).

Cryopreservation component
Serum-free cells, serum-free stem cells and MSC cryopreservation

Storage Conditions
Store below 4 °C. The quality guarantee period is 3 years from the date of production of the product. When the frozen solution plan is not used for more than 3 months, store it as cold as possible. In order to avoid the degradation of the quality of the frozen solution and the performance degradation caused by repeated freeze-thaw cycles, it is recommended to dispense the 100 ml product into small vials and store them in the refrigerator below 4 °C or -20 °C.

Instructions
Cell cryopreservation method
Choosing to cryopreserved cells in the logarithmic growth phase helps to increase the survival rate of resuscitated cells.
1. Collect suspended cells or adherent cells in a test tube according to the usual method.
2. Calculate the number of frozen cells required according to the density of the cultured cells and the size of the cryotubes used.
(Reference: 5 × 105 to 5 × 106 cells / ml).
3. Take the amount of cell suspension corresponding to the required number of cells, place in a centrifuge tube, and collect the cultured cells by centrifugation (refer to the centrifugation conditions: 1,000 to 2,000 rpm, 4 ° C, 3 to 5 min). Remove the supernatant from the centrifuge tube.
4. Add an appropriate amount of serum-free cell cryopreservation solution to the centrifuge tube to a cell concentration of 5 x 105 to 5 x 106 cells/ml. Mix slowly and mix well to make a cell mixture.
5. Dispense the cell mixture in the centrifuge tube into a cryopreservation tube that has been labeled as complete.
6. Directly store the cryotube containing the cell mixture in a -80 ° C refrigerator and store it for a long time.
7. If the investigator needs liquid nitrogen storage, the fully frozen cryotube (at least one day and night after being placed in a -80 °C refrigerator) can be moved to a -196 °C liquid nitrogen tank.

Cryopreservation cell recovery method
1. Remove the cell cryopreservation tube from the refrigerator and immediately thaw it in a 37 °C shaking water bath.
2. After the cell mixture in the cryotube is completely melted, immediately add 1 ml of cell culture medium to the cells in the cryotube, and then transfer the cell mixture from the cryotube to the test tube containing 9 ml of the cell culture medium. ,well mixed.
3. Centrifuge the cultured cells (refer to the centrifugation conditions: 1,000 to 2,000 rpm, 4 ° C, 3 to 5 min), and remove the supernatant.
4. Wash the cells and wash off the residual cryopreservation solution.
5. Add the appropriate amount of fresh cell culture medium and slowly mix the cell mixture with a pipette. After appropriate dilution, the cell mixture is transferred to a previously prepared culture flask.
6. After microscopy, the researchers can perform cell culture according to their respective methods and needs.

Resuscitation cell survival rate

● It is best to cryopreserve cells in logarithmic growth phase.
● Control the digestion time of trypsin, remember that excessive digestion will affect the effect of cell resuscitation.
● During the process of resuspending the cells, be sure to gently so as not to damage the cells, but also must be fully resuspended so that the cells do not clump when they recover.
● Please observe the sterility of cell operation

Hydroxides

Calcium Hydroxide,Slaked Lime,Hydrated Lime,Food Grade Calcium Hydroxide

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