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Make Human Umbilical Vein Endothelial Cells from Cords
Published: Apr 20, 2012 DOI: 10.21769/BioProtoc.204 Views: 13552
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Abstract
Human umbilical vein endothelial cells (HUVEC) can be isolated from normal human umbilical veins, and are responsive to cytokine stimulation by expressing cell adhesion molecules. These cell systems are commonly used for physiological and pharmacological investigations, such as macromolecule transport, blood coagulation, and angiogenesis. This protocol describes the generation of HUVECs.
Materials and Reagents
- Collagenase I (Life Technologies, InvitrogenTM, catalog number: 17100-017 )
- Endothelial cell growth supplement (ECGS) (Sigma-Aldrich, catalog number: E2759 )
- M199 medium
- Fetal calf serum (FCS)
- Phosphate buffered saline (PBS)
- Pen/Strep
- Sodium pyruvate
- Antibiotic
- Ethanol
- Collagenase solution
- Culture medium (see Recipes)
Equipment
- Needle
- Clamps
- Syringes
- Syringe filter
- Parafilm
- Heparin
- Culture hood
- Sterilized beaker
- 50 ml tube
Procedure
Category
Cell Biology > Cell isolation and culture > Cell isolation
Developmental Biology > Cell growth and fate > Umbilical cord
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