CryoDefend-Stem Cells (5 x 10 mL)

For defined, protein-free cryopreservation
Catalog # Availability Size / Price Qty
CCM018
Product Details
Procedure
FAQs
Supplemental Products
Reviews

CryoDefend-Stem Cells (5 x 10 mL) Summary

Kit Summary

Media for defined, protein-free cryopreservation of stem cells.

Key Benefits

  • Reduce experimental variability by using fully defined media
  • Supports post-thaw cell viability better than conventional freezing media
  • Optimized for cryopreservation of stem cells
  • Suitable for all stem cell types
 

 

Why use fully defined, protein-free media to cryopreserve stem cells?

Cryopreservation is essential for the banking of valuable stem cell cultures. The viability of the recovered cells depends on cell density, the type of cryoprotectant used, the method of cooling, and the amount of the cryoprotectant that remains in the culture after the cells have been thawed.

Additional considerations for stem cell researchers include the effect of the cryopreservative on stem cell potency and the presence of any undefined factors which could limit t he use of recovered cells in preclinical and clinical studies. These additional considerations emphasize the importance of using cryopreservation media that has been specifically optimized for the cryopreservation of stem cells.

R&D Systems offers CryoDefend® Stem Cells Media, a defined, protein-free media, which has been optimized for stem cell cryopreservation.  This specially formulated media increases the recovery and viability of healthy undifferentiated cells compared to conventional freezing media.

 

 

Kit Contents

Components

  • 5 x 10 mL vials of CryoDefend® Stem Cells Media

Stability and Storage

Upon receipt, the CryoDefend® Stem Cells Media should be stored at = -20 °C in a manual defrost freezer. The media can be thawed at 2 °C to 8 °C or at room temperature. Thawed media can be aliquoted and stored at = -20 °C in a manual defrost freezer for up to 3 months. Thaw a fresh aliquot for each use. Avoid repeated freeze-thaw cycles.

 

Embryonic stem (ES) cells are pluripotent stem cells derived from the inner cell mass of pre-implantation embryos. Induced pluripotent stem (iPS) cells can be generated by somatic cell reprogramming following the exogenous expression of specific transcription factors (Oct-3/4, KLF4, SOX2, and c-Myc). These cell types are capable of unlimited, undifferentiated proliferation in vitro and still maintain the capacity to differentiate into a wide variety of somatic cells. In this capacity, pluripotent stem cells have widespread clinical potential for the treatments of heart disease, diabetes, spinal cord injury, and a variety of neurodegenerative disorders.

R&D Systems offers a wide range of products to support pluripotent stem cell culture and differentiation. Mouse embryonic fibroblasts may be used to maintain and expand pluripotent stem cells in an undifferentiated state. We also offer defined culture media, which are specifically optimized for use with human or rodent pluripotent stem cells. In addition, R&D Systems offers a variety of products to assess differentiation status and identify specific stem cell types of interest, including panels of marker antibodies, primer pairs, multi-color flow cytometry kits, and specialized verification kits.

Specifications

Shipping Conditions
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Species
Human

Product Datasheets

Assay Procedure

Refer to the product datasheet for complete product details.

  • Transfer detached cells to a conical tube and centrifuge
  • Remove supernatant and resuspend in CryoDefend® Stem Cells Media
  • Transfer cells to a cryovial and freeze at -80 °C overnight
  • Transfer the frozen cryovial to liquid nitrogen
 

 

Reagents Provided

Reagents supplied in the CryoDefend® Stem Cells Media (Catalog # CCM018):

  • 5 x 10 mL vials of CryoDefend® Stem Cells Media

 

Other Supplies Required

Reagents

  • Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005)

Materials

  • Stem cells
  • Cryovials
  • 15 mL centrifuge tubes
  • Serological pipettes
  • Pipette and pipette tips

Equipment

  • 37 °C and 5% CO2 incubator
  • Centrifuge

 

Procedure Overview

Cryopreservation of Stem Cells

Thaw CryoDefend® Stem Cells Media.

Thaw CryoDefend Stem Cells Media.

Detach the cells from the cell culture dish.

Detach the cells from the cell culture dish.

Transfer the cells to a 15 mL conical tube.

Centrifuge at 200 x g for 5 minutes.

Remove and discard the supernatant.

Resuspend the cells in CryoDefend® Stem Cells Media at 0.5-1.0 x 106 cells/mL.

Transfer the cells to a 15 mL conical tube.

Transfer the cells to a cryovial.

Freeze the cryovial at -80 °C overnight.

Transfer the frozen cryovial to liquid nitrogen for storage.

Transfer the cells to a cryovial.
 

Thawing of Cryopreserved Stem Cells

Warm appropriate cell culture media to 37 °C.

Warm appropriate cell culture media to 37  C.

Add pre-warmed culture media to the cryovial containing cryopreserved cells.

Pipette up and down and as cells thaw, transfer the thawed cells to a 15 mL conical tube containing pre-warmed expansion media.

Add pre-warmed culture media to the cryovial containing cryopreserved cells.

Centrifuge at 200 x g for 5 minutes.

Resuspend the cells in pre-warmed expansion media.

Plate the stem cells at the desired density.

Centrifuge at 200 x g for 5 minutes.

FAQs

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