TC Protector

TC protector is a cell cryoprotective agent that does not contain any animal-derived components nor proteins and is chemically defined. It can be used in cells requiring serum addition, those without serum, and cells without proteins.

Characteristics
· It does not contain animal-derived components at all and is chemically defined.
· It does not contain any serum or protein.
· It is stored in a refrigerator and can be used immediately after storage.
· It is less viscous and usability is excellent.
· It does not require special equipment such as Programed Freezer. It can be directly stored at −80ºC

Freezing method
1. In case of adhesive cells, media should be changed at 80% confluence and be frozen on the next day.
In case of floating cells, media change (or media supplementation) is conducted and should be frozen on the next day.
*In order to freeze cells in a good condition, it is important to use cells from the logarithmic growth phase.
*Cells that have reached confluence or overgrown result in a decreased survival rate after freezing.
2. Cells are harvested according to the standard procedure and are centrifuged at 1,500 rpm for 1 minute.
3. It is diluted with TC Protector to achieve 5×10^5～1×10^7 cell/mL.
*Cell suspension is maintained in ice water when cell counts are measured.
*Since the appropriate cell number varies depending on cells, a pilot study must be conducted in advance. In general, 1×106 cells/mL is the target cell count.
4. Cells are stored at −80ºC in a Deep Freezer and are transferred into liquid nitrogen on the next day.

Thawing method
· Ampules are placed in a warm water bath at 37ºC, and it is quickly thawed until it forms small ice mass.
· Cell suspension is collected into a spitz tube and about 10× media is added. Then, it is centrifuged at 1,000–1,500 rpm for 1–2 minutes.
· Cell pellet is checked and supernatant is discarded. Cells are seeded in accordance with the standard method.

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