Cat. No. VPL-A1
Size 20 ml
NIH NSN# 655000L129024
- Prepare liquid N2 in the clean bench.
- Centrifuge suspended media of ES/iPS cell colonies and pellet them by elimination of supernatant at 4 ~ 10 °C.
- Resuspend the cell pellet with 200 L of StemCell Keep.
- Pipette the cell suspension into a cryotube and immediately put it to liquid N2 directly. (If the successful vitrification would be achieved, the solution should be kept transparent after freezing.)
- Transfer the cryotube into a liquid nitrogen chamber (-196 °C).
- Keep the vials to be thawed in the liquid N2 in the Dewar vessel.
- Add the pre-warmed (37 ) media (1mL) directly to the vial immediately after taking out from liquid N2.
- Thaw the solution by gentle pipetting and add into the excess amount of medium. Centrifuge and plated the colonies on the feeder cell layer in a culture dish and culture in an incubator.
contamination is all negative.
(2) Endotoxin < 0.5 EU/mL
Very old method of cryopreservation was “Slow Programmed Freezing” using by 10% DMSO contained solution. However, big ice crystal is formed by this method and this crystal causes very poor cell viability (0.1-1%) for primate ES and iPS cells. Improved method is “vitrification” using by DAP213 (DMSO 2M, Acetamide 1M, Propylene glycol 3M).
Vitrification is the new method for preventing ice crystal formation by rapid freezing (＜15 sec) in LN2. However, this method requires rapid thawing to prevent recrystallization and experienced operators. It is known that DMSO has influence on OCT-4 expression and differentiation, Acetamide is identified as carcinogens. Moreover, DAP213 is high osmolarity solution – it means this solution has high toxicity. Therefore, development of low Toxicity, easy to use vitrification solution had been expected.StemCell Keep is the solution that overcomes these disadvantages of DAP213!
Solution freezed by Liquid Nitrogen is heated by 50℃/min.
StemCell Keep does not have recrystallization during temperature rising.
- Animal-derived Protein-free, Serum-free and DMSO-free formulation. No risk of differentiation effect by DMSO.
- Maintain ES and iPS cells with colony formation by vitrification.
- Maintain stem cell pluripotency after thawing.
- High cytoprotection effect and vitrification technology by new original cryoprotectant material is allowed to cryopreserve ES and iPS cell colonies.
- Sufficient for 100 vials.
- Free of bacteria, fungi and mycoplasma contamination.
- Long shelf life. The product is stable for 2 years at 4C after the date of manufacture.
- Free Trial Sample is available! (Please contact us)