TC-Protector is a chemically defined cell cryo-protective agent that does not contain any animal-derived components or proteins. It allows direct freezing of variety of cell lines at -80°C. Cells cryo-preserved with TC-protector have shown a high cell viability and durability rate on thawing.
It can be used in cells requiring serum addition, cells without serum, and cells without proteins. It is suitable for both cell line and normal cells, as well as ES cells, adult stem cells, and human iPS cells. It is not viscous and does not form bubbles, making it excellent for use in these contexts.
100 mL x 1 or 10 mL x 10
Protect from light and storage at 4°C, avoid freezing prior to use
- Free of animal-derived components
- Does not contain any serum or protein
- Low viscosity and easy to use
- Can be stored in a refrigerator and used immediately
- Does not require special equipment to freeze cells
Cryo-preserve cells directly without using programmed freezer at -80°C.
Instructions for Use
1. Cell Preparation:
- For adherent cells, media should be changed at 80% confluence one day before cryopreservation. For the suspension cultured cells, change or supplement media one day before the cryopreservation.
- It is important to preserve cells during the logarithmic growth phase for the best result.
- Cryo-preserving confluent or overgrown cells will result in a decreased viability rate on thawing.
2. Harvest the cells by centrifuging cells in medium at 1,500 rpm for 1 minute.
3. Remove supernatant and re-suspend cells in TC-Protector at a concentration of between 5x106 and 1x107 cells/mL.
Notes: Cell suspension should be kept on ice while preparing an appropriate (adequate) cell density. Appropriate cell number varies depending on the type of cell. Preliminary studies are required for determining the optimum cell density. In general, 1×106 cells/mL is a recommended starting density.
4. Freeze the cells in a deep freezer at -80 ºC. Transfer to liquid nitrogen if preferred on the following day.
1. Thaw frozen vials rapidly in a warm water bath at 37ºC until it becomes a small ice mass.
2. Transfer the cell suspension to a conical centrifuge tube and dilute by adding about x10 media. Centrifuge at 1,000～1,500 rpm for 1～2 minutes.
3. Discard the supernatant after verifying cell pellet at the bottom of the tube.
4. Seed cells in accordance with the standard method.
Miyagi-Shiohira, C. et al. Evaluation of Serum-Free, Xeno-Free Cryopreservation Solutions for Human Adipose-Derived Mesenchymal Stem Cells. Cell Med 9, 15–20 (2016).https://doi.org/10.3727/215517916X693122
Takebe, T. et al. Transient vascularization of transplanted human adult-derived progenitors promotes self-organizing cartilage. J Clin Invest 124, 4325–4334 (2014). https://doi.org/10.1172/JCI76443
Miwa, H., Hashimoto, Y., Tensho, K., Wakitani, S. & Takagi, M. Xeno-free proliferation of human bone marrow mesenchymal stem cells. Cytotechnology 64, 301–308 (2012).https://doi.org/10.1007/s10616-011-9400-7
Q: How long can I keep cells at - 80ºC with specially designed freezing media?
A: KAC recommends not to exceed a few months in a deep freezer, due to the temperature fluctuation caused by opening and closing the freezer door. If you want to cryopreserve for more than one month, using liquid nitrogen is recommended. Cells can be preserved indefinitely by storing in liquid nitrogen.
Q: Does TC-Protector contain calcium and magnesium?
A: Yes. However, the actual concentration is proprietary information.
Q: What is the composition of TC-Protector?
A: It is proprietary information. We can disclose only if we have a non-disclosure agreement (NDA).
Q: Does TC-Protector contain any components of animal origin?
A: There are no components of animal origin.
Q: What is the ratio to mix TC-Protector and media when cryopreserving cells?
A: TC-Protector is provided in a ready-to-use form. Do not mix with media. Simply suspend cells in TC-protector, verify the cells are not aggregated then place the vial in deep freezer.