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Endothelial Cell Growth Media

Endothelial Cell Base Media plus Supplement
Improved Proliferation of Primary Human Dermal Lymphatic Endothelial Cells_CCM027
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Endothelial Cell Growth Media
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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
 

 

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

Warm Complete Endothelial Cell Growth Media to 37 °C.

Determine the size and number of flasks needed for plating.

Warm Complete Endothelial Cell Growth Media to 37 °C.

Warm the frozen vial of HUVECs until just thawed.

Warm the frozen vial of HUVECs until just thawed.

Transfer cells immediately and gently into a 15 mL centrifuge tube containing > 5 mL of prewarmed Complete Endothelial Cell Growth Media.

Centrifuge at 200 x g for 5 minutes.

Transfer cells immediately and gently into a 15 mL centrifuge tube > 5 mL of prewarmed Complete Endothelial Cell Growth Media.

Resuspend the pellet in prewarmed Complete Endothelial Cell Growth Media

Transfer HUVEC suspension into a T75 flask(s).

Resuspend the pellet in prewarmed Complete Endothelial Cell Growth Media

Change media the day following thaw.

Monitor cells daily and change media every other day.

Passage the cells at 80-90% confluency.

Change media the day following thaw.
 

Subculturing of HUVECs

Warm Complete Endothelial Cell Growth Media to 37 °C.

Warm Complete Endothelial Cell Growth Media to 37 °C.

Remove and discard the media from the flasks.

Rinse cells once with sterile 1X PBS.

Digest cells with TrypLE Express for 1-2 minutes at 37 °C.

Remove and discard the media from the flasks.

Tap the side of the flask to aid the detachment of the cells.

Transfer the cell suspension to a 15 mL centrifuge tube containing 8 mL of prewarmed Complete Endothelial Cell Growth

Centrifuge at 200 x g for 5 minutes.

Tap the side of the flask to aid the detachment of the cells.

Plate cells (approximately 6.7 x 103 cells/cm2).

Change media the day following thaw.

Monitor cells daily and change media every other day.

Passage the cells at 80-90% confluency.

Plate cells (approximately 6.7 x 103 cells/cm2)

Citations (1)

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Customer Reviews (6)

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  • Endothelial Cell Growth Media
    Name: Anonymous
    Verified Customer | Posted 11/05/2018
    Endothelial Cell Growth Media Serum CCM027
  • Endothelial Cell Growth Media
    Name: Anonymous
    Verified Customer | Posted 07/11/2018
  • Endothelial Cell Growth Media
    Name: Anonymous
    Verified Customer | Posted 04/03/2018
  • Endothelial Cell Growth Media
    Name: Anonymous
    Verified Customer | Posted 02/03/2018
    Im working with HUVEC cells for the first time. I was very confused about the medium, so I ordered this medium. It was delivered on time and had needed growth factor supplements for HUVEC. I will highly recommend this medium.
    Endothelial Cell Growth Media Serum CCM027
  • Endothelial Cell Growth Media
    Name: Anonymous
    Verified Customer | Posted 01/11/2018
    Our primary retina endothelial cells grow fast and nice with this media.
    Endothelial Cell Growth Media Serum CCM027
  • Endothelial Cell Growth Media
    Name: Anonymous
    Verified Customer | Posted 12/27/2017
    We culture primary Primary Human Retinal Microvascular Endothelial Cells. The cell grow very well by meida and 50X supplement.
    Endothelial Cell Growth Media Serum CCM027

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Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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