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CryoDefend-Stem Cells (5 x 10 mL)

Bio-Techne includes R&D Systems | Catalog # CCM018

For defined, protein-free cryopreservation

Key Product Details

Human Embryonic Stem Cell Viability Following Cryopreservation in CryoDefend® Stem Cells Media or Cryopreservation Media from Two Competitors. 
(6)
Morphology of Human Embryonic Stem Cells Following Cryopreservation in CryoDefend® Stem Cells Media, Conventional Freezing Media, or Freezing Media from a Competitor.
Expression of Pluripotency Markers in BG01V Human Embryonic Stem Cells Frozen in CryoDefend® Stem Cells Media or Cryopreservation Media from Two Different Competitors.
Human Embryonic Stem Cells Maintain Pluripotency Following Cryopreservation in CryoDefend® Stem Cells Media.
Recovery and Marker Expression of Rat Mesenchymal Stem Cells Cryopreserved in CryoDefend® Stem Cells Media or Conventional Freezing Media. 
Rat Cortical Stem Cell Viability Following Cryopreservation in Control Cyropreservation Media or CryoDefend® Stem Cells Media. 

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.

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Product Documents for CryoDefend-Stem Cells (5 x 10 mL)

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