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Primary interstitial (fibroblast) cells and endothelial cells were isolated from porcine aortic valve leaflets, as previously described [30], [31], and cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS); and in Medium 199 supplemented with 10% FBS, respectively.
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The AVL and MVL calcifications were graded as 0 (absent), 1 (mild, meaning only one leaflet affected), or 2 (severe, 2 or 3 leaflets affected), as previously described.
The heterogeneity in organelles and membrane leaflets is, as previously mentioned, critical in modulating the localization of membrane-bound proteins and their functional properties.
Analytical methods were as previously described [33].
VICs were isolated by sequential collagenase digestion of porcine aortic leaflets as described previously (24).
In particular, PE redistributes to the exoplasmic leaflet as well.
Severe AR was induced by retrograde puncture of the aortic valve leaflets under echocardiographic guidance as previously described [ 16– 16].
Data from open questions were grouped according to whether they referred to leaflet content or structure and analysed as previously.
As previously reported13,17.
as previously published36.
As previously stated: messy.
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