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Endothelial Cell Growth Media
R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
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Endothelial Cell Growth Media Supports Cell Proliferation of Human Umbilical Cord Endothelial Cells (HUVECs). One T25 flask of HUVECs (0.17 x 106cells) were expanded over 10 passages using Complete Endothelial Cell Growth Media. Cells were passaged every 4-6 days. The histogram shows the cumulative number of HUVECs expanded over 10 passages if all cells were replated at each passage. |
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Human Umbilical Cord Endothelial Cells (HUVECs) Express Endothelial Cell Markers CD31, KDR, and vWF. HUVECs were expanded in Complete Endothelial Cell Growth Media for 5 passages prior to staining for marker expression. (A,B) Flow cytometry analysis of endothelial cell marker expression. Filled histograms indicate cells stained with PE-conjugated Mouse Anti-CD31 (A; R&D Systems®, Catalog # FAB3567P) or PE-conjugated Mouse Anti-KDR (B; R&D Systems®, Catalog # FAB357P). Open histograms in A and B indicate staining with PE-conjugated Mouse IgG1 isotype control (R&D Systems®, Catalog # IC002P). (C) Immunocytochemistry analysis of endothelial cell marker expression. Cells were double stained with Mouse Anti-CD31 (red; R&D Systems®, Catalog # BBA7) and Rabbit Anti-Von Willebrand Factor (vWF) (green; Novus Biologicals®, Catalog # NB600-586). The nuclei were counterstained with DAPI (blue) |
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Increased HUVEC Survival and CD31 Expression Compared to Competitor Endothelial Cell Media. HUVECs were expanded for 14 passages in either R&D Systems Endothelial Cell Growth Media (Catalog # CCM027) or competitor endothelial cell media. Cells were stained for CD31 expression using the Mouse Anti-Human CD31 Antibody (yellow; R&D Systems; Catalog # BBA7) and counterstained with DAPI (Catalog # 5748).A) Representative images of CD31 expression in p14 HUVECs grown in R&D Systems Media or competitor media. There is a greater density of cells and a higher expression of CD31 in HUVECs grown in R&D Systems Media.B) Histogram showing that at p14, there is a higher percentage of surviving cells that express CD31 in HUVECs grown in R&D Systems Media compared to competitor. |
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Improved Yield of Murine CD31+PDGFa- Lung Cells using Endothelial Cell Growth Media A single cell suspension of mouse lung cells was prepared and seeded onto gelatin coated plates in 3 different media, R&D Systems®Endothelial Cell Growth Media (Catalog # CCM027), competitor media, or 20% FBS/DMEM. After one week in culture, cells were analyzed for expression of CD31 (APC) and PDGFRα (PE). The percentage of CD31+PDGFRα- cells obtained from R&D Systems Endothelial Cell Growth Media was superior (13.2%) compared to competitor (3.5%) and 20% FBS/DMEM (4.5%) media. Data courtesy of the laboratory of Dr. Michael Kyba at the University of Minnesota. |
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Improved Proliferation of Primary Human Dermal Lymphatic Endothelial Cells. Human dermal lymphatic endothelial cells (HDLECs) were cultured in either Endothelial Cell Growth Media (R&D Systems, Catalog # CCM027) or a competitor endothelial cell media. After 48 hours, HDLEC proliferation was assessed using a MTT proliferation assay. HDLECs cultured in R&D Systems Endothelial Cell Growth Medium showed a 24% increase in proliferation compared to competitor media. The P-value was calculated using an unpaired t-test. Data provided courtesy of the laboratory of Dr. Constantinos Mikelis at Texas Tech University Health Sciences Center. |
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Improved Yield of Murine CD31+PDGF alpha - Lung Cells using Endothelial Cell Growth Media. A single cell suspension of mouse lung cells was prepared and seeded onto gelatin coated plates in 3 different media, R&D Systems Endothelial Cell Growth Media (Catalog # CCM027), competitor media, or 20% FBS/DMEM. After one week in culture, cells were analyzed for expression of CD31 (APC) and PDGFR alpha (PE). The percentage of CD31+PDGFR alpha - cells obtained from R&D Systems Endothelial Cell Growth Media was superior (13.2%) compared to competitor (3.5%) and 20% FBS/DMEM (4.5%) media. Data courtesy of the laboratory of Dr. Michael Kyba at the University of Minnesota. |
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 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.
Resuspend the pellet in prewarmed Complete Endothelial Cell Growth Media
Transfer HUVEC suspension into a T75 flask(s).
Change media the day following thaw.
Monitor cells daily and change media every other day.
Passage the cells at 80-90% confluency.
Subculturing of HUVECs
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.
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.
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.
Citations for Endothelial Cell Growth Media (5)
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Citations are publications that use Bio-Techne products. Selected citations for Endothelial Cell Growth Media include:
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Species: Human
Sample Types: Whole Cells
Applications: BioassayDB AbuSamra et al. (2021), Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins Cells, 10(2).
PMID: 33573183 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayTD Saccon et al. (2021), Circulating microRNA profile in humans and mice with congenital GH deficiency Aging Cell, 0(0):e13420.
PMID: 34118183 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayA Lee et al. (2021), Amino Acid Nanofibers Improve Glycemia and Confer Cognitive Therapeutic Efficacy to Bound Insulin Pharmaceutics, 14(1).
PMID: 35056977 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayW Zhou et al. (2020), Mitofusin 2 regulates neutrophil adhesive migration and the actin cytoskeleton J. Cell. Sci., 133(17).
PMID: 32788232 -
Species: Human
Sample Types: Whole Cells
Wu Z et al. (2017), Imaging specific cellular glycan structures using glycosyltransferases via click chemistry. Glycobiology, 0(0).
PMID: 29186441
There are no citations that match your criteria.
Customer Reviews for Endothelial Cell Growth Media (8)
5 out of 5
8 customer ratings
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Verified Customer | Posted 03/01/2022Tube assay using culture media with supplemented growth factors
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Verified Customer | Posted 03/01/2022Tube assay using CCM027 culture media
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Verified Customer | Posted 11/05/2018
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Verified Customer | Posted 07/11/2018
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Verified Customer | Posted 04/03/2018
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Verified Customer | Posted 02/03/2018Im 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.
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Verified Customer | Posted 01/11/2018Our primary retina endothelial cells grow fast and nice with this media.
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Verified Customer | Posted 12/27/2017We culture primary Primary Human Retinal Microvascular Endothelial Cells. The cell grow very well by meida and 50X supplement.
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