Cryoderm - glycerin-based cryopreservation medium for cells, intended for use in surgery and ophthalmology when creating stocks of viable skin and cornea transplants, as well as for cryopreservation of cells in biotechnology.
This product is intended for research purposes.
Instructions for use
1. Take a culture with a monolayer density of 70-80% (for cells of attached cultures), or 50-70% of suspension density (for cells of suspension cultures).
2. Remove the cells of the attached cultures from the substrate using dissociating 0.25% trypsin solution for 5 minutes at 37oC (pre-rinse the monolayer of cells with 5 ml of 0.02% Versin solution). The cells of suspension cultures are pipetted and collected in a centrifuge tube.
3. Inactivate trypsin with a medium containing 10% serum.
4. Precipitate the cells by centrifugation 1000 rpm for 5 minutes. Remove the supernatant without affecting the cellular sediment.
5. Resuspend cells in the "Cryoderm" medium at the rate of 1.2-1.5 million/ml.
6. Pour the prepared suspension into cryoprobes, seal the frozen samples hermetically.
7. Freezing is carried out in stages - first the cryoprobes are cooled at temperature of 2 ± 8oC for 5 minutes, and then they are placed in the freezer at -18 ± 2oC for 24 hours.
Defrosting of cells
* Completely defrost the contents of cryoprobes at 37oC.
* Gently resuspend the cell sediment in a cryoprobe and put the entire volume of sample into a tube with 10 ml of appropriate culture medium.
Thoroughly disperse the cells and centrifugate at 1000 rpm for 2 minutes. Drain the infusion fluid, and resuspend the resulting precipitate in 1 ml of a nutrient medium containing 10% bovine blood serum.
* Before the experiment, make 1-2 passages of culture under appropriate conditions.
• For maximum removal of cryoprotectors, it is recommended to completely replace the medium 1 day after the start of cell culture.
* If necessary, before the passage, evaluate the number of viable cells in the seed material by staining with trypan blue.
Shelf life: 1 year
Storage and transportation conditions:
Store at temperature of 2-8 ° C in a place protected from light. Transportation by all types of covered transport at temperature of 2-8 oC.