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Endothelial Cell Growth Media Best Seller

R&D Systems, part of Bio-Techne | Catalog # CCM027

Endothelial Cell Base Media plus Supplement
R&D Systems, part of Bio-Techne

Key Product Details

Endothelial Cell Growth Media Supports Cell Proliferation of Human Umbilical Cord Endothelial Cells (HUVECs).
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CCM027

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, Complete Endothelial Cell Growth Media is prepared using the following procedure:

  • Add Endothelial Growth Media Supplement to Endothelial Cell Growth Base Media
  • Store completed media at 2 - 8 °C
    View Graphic Procedure

 

Reagents Provided

Reagents provided with the Endothelial Cell Growth Media (Catalog # CCM027):

  • Endothelial Cell Growth Base Media Human
  • Endothelial Cell Growth Supplement (50X)

    Note: The components of this list require different storage/shipping temperatures and will arrive in separate packaging.

 

Other Supplies Required

Reagents

  • HUVECs
  • Penicillin-Streptomycin (100X), optional
  • TrypLE Express
  • Phosphate-Buffered Saline (PBS) (Tocris; Catalog # 3156)

Materials

  • 75 cm2 tissue culture (T75) flasks
  • 15 mL centrifuge tubes
  • Serological pipettes
  • Pipette and pipette tips

Equipment

  • 37°C and 5% CO2 humidified incubator
  • Centrifuge (low speed clinical or equivalent)
  • Hemocytometer
  • Inverted microscope
  • Water bath

 

Procedure Overview

Reagent Preparation

Endothelial Cell Growth Supplement - Thaw the Endothelial Cell Growth Supplement (50X) at 2-8 °C or room temperature.

Complete Endothelial Cell Growth Media - Add 5 mL of Endothelial Cell Growth Supplement to 250 mL of Endothelial Cell Growth Base Media. Add Penicillin-Streptomycin at a 1:100 dilution. Store under sterile conditions at 2-8 °C for up to 2 weeks.

Note: If Penicillin-Streptomycin is not needed for the experiment, it can be omitted.

Culturing of HUVECs

  1. Warm Complete Endothelial Cell Growth Media to 37 °C.
  2. Determine the size and number of flasks needed for plating.
  3. Warm the frozen vial of HUVECs until just thawed.
  4. Transfer cells immediately and gently into a 15 mL centrifuge tube containing > 5 mL of prewarmed Complete Endothelial Cell Growth Media.
  5. Centrifuge at 200 x g for 5 minutes.
  6. Resuspend the pellet in prewarmed Complete Endothelial Cell Growth Media.
  7. Transfer HUVEC suspension into a T75 flask(s).
  8. Change media the day following thaw.
  9. Monitor cells daily and change media every other day.
  10. Passage the cells at 80-90% confluency.

Subculturing of HUVECs

  1. Warm Complete Endothelial Cell Growth Media to 37 °C.
  2. Remove and discard the media from the flasks.
  3. Rinse cells once with sterile 1X PBS.
  4. Digest cells with TrypLE Express for 1-2 minutes at 37 °C.
  5. Tap the side of the flask to aid the detachment of the cells.
  6. Transfer the cell suspension to a 15 mL centrifuge tube containing 8 mL of prewarmed Complete Endothelial Cell Growth
  7. Centrifuge at 200 x g for 5 minutes.
  8. Plate cells (approximately 6.7 x 103 cells/cm2).
  9. Change media the day following thaw.
  10. Monitor cells daily and change media every other day.
  11. Passage the cells at 80-90% confluency.
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Product Documents for Endothelial Cell Growth Media

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.