Serum-free cell cryopreservation solution
Packing specification
Product Numbers:CE70010、CE70020、CE70040、CE70080、CE70100T
Specification: 2*10ml、4*10ml、8*10ml、100ml
Storage conditions
Store at 4oC, valid for one year; Store at -20oC, valid for two years;
feature of product
Ready to use, easy to save time;
No serum composition, clear chemical composition;
Quick freezing, can be placed directly in the refrigerator at -80 ℃;
High recovery rate and effective protection of cells;
Freezing operation steps
1. Collect suspended cells or adherent cells in the logarithmic growth phase;
(※ Cryopreservation of cells in logarithmic proliferation phase is the most important point to ensure a good cell survival rate.)
2. Use some cells to stain with trypan blue and count the number of cells;
3. Transfer the cells that need to be frozen into a centrifuge tube, 1000rmp, centrifuge for 5 minutes to collect the cultured cell pellet, and completely discard the supernatant in the centrifuge tube;
4. Add an appropriate amount of cell cryopreservation solution to the centrifuge tube so that the cell concentration is about 5x105-1x107 / ml and gently mix the cells to make a cell mixture suspension;
5. Dispense the cell suspension mixed with cryopreservation solution into 1ml or 1.5ml cryopreservation tubes;
(※ The following operation steps should be completed as quickly as possible. In addition, the frozen solution should be stored as soon as possible after it is taken out of the refrigerator.)
6. Without pre-chilled cell suspension, store directly at -80℃;
7. If the cell suspension is to be stored in liquid nitrogen, first store it at -80℃ for 12 hours (more than one night), and then move to liquid nitrogen after the state is stable.
Frozen cell recovery steps
1. Thaw the cells in a 37℃thermostat (or water bath) and other equipment to confirm that the cell fluid is completely thawed.
(※ This operation needs to be performed quickly. Because the cells are extremely vulnerable to damage during the thawing process)
2. After the cell mixture in the cryotube is completely thawed, immediately add 1ml of cell culture medium to the cryotube and mix with the cells.
Transfer the mixed solution to a centrifuge tube containing about 5ml of the cell culture medium, 1000rmp, centrifuge for 5 minutes to collect the frozen
Store the cell pellet and remove the supernatant (be careful when handling, do not remove the cell pellet).
3. Add an appropriate amount of fresh cell culture medium, slowly add to the cell pellet using a pipette, mix gently, and transfer the cell mixture to the culture vessel prepared in advance.
Sterility testing
Endotoxin: chromogenic substrate method
Mycoplasma: fluorescent antibody method
Fungi and bacteria: according to the US Pharmacopoeia
Verified cell information
Cell name | Save time | Recovery efficiency |
k562 | 12months | 98% |
RAW264.7 | 12months | 95% |
HEK-293 | 7months | 98% |
Hela | 12months | 95% |
A549 | 12months | 95% |
NCI-H460 | 12months | 89% |
CHO | 12months | 90% |
MCF-7 | 12months | 90% |
HepG2 | 12months | 95% |
C2C12 | 12months | 95% |
CHO | 12months | 95% |
COS7 | 12months | 95% |
DU145 | 12months | 95% |
MDCK | 12months | 95% |
NIH-3T3 | 12months | 90% |
MSC | 12months | 90% |
Precautions
1. Store the low-temperature reagents at room temperature for 5 minutes before use.
2. Please choose to freeze the logarithmic growth phase cells;
3. After adding the cryopreservation solution to the cells, please put them into the -80oC refrigerator for storage as soon as possible;
4. The frozen cells of this product can be stored in the refrigerator at -80oC for more than 5 years;
5. If you need to freeze the cells for a long time, please transfer to a liquid nitrogen tank for storage;
6. The frozen storage solution contains DMSO ingredients. For your safety and health, please wear laboratory clothes and disposable gloves.
For scientific research use only.