Primary Huvec Cell Culture

Add 2 ml of the cell suspension in each fibronectin coated petri dish cultures are placed in a humidified 95 air and 5 co2 atmosphere at 37 c.

Primary huvec cell culture.

Warm complete growth medium to 37 c prior to use with the cells. All cells test negative for mycoplasma bacteria yeast and fungi. Aspirate the medium from a flask of primary huvec isolated from one or more umbilical cords at or near confluence see refs. The culture medium is changed every two days and confluency is typically achieved in 6 8 days with cobblestone.

Product overview lonza s cryopreserved huvecs are shipped as frozen primaries. How do other people freeze down and thaw huvec. Pipette 20 ml of endothelial cell growth medium to a t 75 flask. Coating cell culture dishes both flasks for expansion and or experimental dishes plates wells with 0 1 gelatin is fine.

Human umbilical vein endothelial cells huvec provide a classic model system to study many aspects of endothelial function and disease such as normal abnormal and tumor associated angiogenesis oxidative stress hypoxia and inflammation related pathways in endothelia under normal and pathological conditions cardiovascular related complications associated with various diseases mode of. The following day non adherent cells are removed by changing the culture medium. Wash the monolayer with 5 ml of hbss volumes are given for a 75 cm 2 flask. They are guaranteed to express cd31 105 and survive in culture through 15 population doublings except huvec xl tm guaranteed through 5 pd.

Passage normal huvec cells when culture has reached approximately 80 confluence. Lonza offers a comprehensive range of normal human primary endothelial cells and supporting media for cardiac aortic coronary iliac artery pulmonary artery cardiac microvascular. Performance testedat least 16 population doublings when cultured in medium 200 supplemented with low serum growth supplement lsgs guarant. To have good huvec monolayer a critical component is ecm.

Preparing cell culture flasks for culturing huvec. Take the endothelial cell growth medium from the refrigerator decontaminate the bottle with 70 alcohol in a sterile hood. Warm both the trypsin edta for primary cells atcc pcs 999 003 and the trypsin neutralizing solution atcc pcs 999 004 to room temperature prior to dissociation. Keep the medium to surface area ratio at 1ml per 5 cm 2.

Use 1 2 as much for 25 cm 2 flasks.