StemCell Keep

Product#: VPL-A1
$200.00
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StemCell Keep

Cat. No. VPL-A1
Size 20 ml
Storage 4°C
NIH NSN# 655000L129024


Description

Let’s preserve your cells “no DMSO and Serum “ condition!
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[ For cryopreservation of Stem Cells = the "StemCell Keep" ]

Protocol
StemCell Keep is a suitable ready-to-use vitrification solution for human ES/iPS cell preservation. Human ES/iPS cells are very sensitive for freezing and thawing damage and their survival rate after cryopreservation by slow cooling is quite low. So vitrification is recommended for preservation of these cell lines. StemCell Keep has been developed for high efficient, low toxic and easy handling vitrification solution without any protein and DMSO which might affect differentiation. 

Usage
1. Freezing of ES/iPS cell colonies suspensions
  1. Prepare liquid N2 in the clean bench.
  2. Centrifuge suspended media of ES/iPS cell colonies and pellet them by elimination of supernatant at 4 ~ 10 °C.
  3. Resuspend the cell pellet with 200 L of StemCell Keep.
  4. 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.)
  5. Transfer the cryotube into a liquid nitrogen chamber (-196 °C).
2.Thawing from preserved state
  1. Keep the vials to be thawed in the liquid N2 in the Dewar vessel.
  2. Add the pre-warmed (37 ) media (1mL) directly to the vial immediately after taking out from liquid N2.
  3. 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.
Expiration
StemCell Keep can be used for 2 years after the manufacture date if stored properly at lower than 10 °C.

Storage
Left-over solution can be stored at 4 °C or –20 °C for 2 years without loosing its functional quality

Quality Control

Bacterial tests:
Bacteria, fungi or mycoplasma
contamination is all negative.
Chemical tests:
(1) pH = 6.7 ~ 8.0 at RT
(2) Endotoxin < 0.5 EU/mL
Memo
Existent problem of preservation media contains DMSO and What is vitrification?

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!

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Solution freezed by Liquid Nitrogen is heated by 50℃/min.
StemCell Keep does not have recrystallization during temperature rising.

Features

  • Animal-derived Protein-free, Serum-free and DMSO-free formulation. No risk of differentiation effect by DMSO.
  • Xeno-freee.
  • 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) 
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Comparison Data
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Outline Video
 

Handling Video
 



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