Permeability Assay for Cerebend Cells

Wild type (WT) and PcdhgC3 knockout (KO) cerebEND cells were grown on gelatin-coated transwells (pore size 0.4 µm, Corning) for 6 days. The permeability assay was performed as previously described (Curtaz et al., 2020 (link)) using 1 µM fluorescein (376 kDa, Sigma-Aldrich) for 1 h with aliquots taken from the basolateral compartment every 20 min. A parallel assay with cell-free transwells was used to calculate the endothelial permeability coefficient (Pe). The Pe of KO cells was normalized to WT cells.

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Gabbert L., Dilling C., Meybohm P, & Burek M. (2020). Deletion of Protocadherin Gamma C3 Induces Phenotypic and Functional Changes in Brain Microvascular Endothelial Cells In Vitro. Frontiers in Pharmacology, 11, 590144.

Publication 2020

gelatin-coated transwells

Assay Cells type Endothelial Fluorescein Gelatin Permeability

Corresponding Organization :

Other organizations : University of Würzburg

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Variable analysis

independent variables

Cell type (Wild type (WT) and PcdhgC3 knockout (KO) cerebEND cells)

dependent variables

Endothelial permeability coefficient (Pe)

control variables

Cells were grown on gelatin-coated transwells (pore size 0.4 µm, Corning) for 6 days
Permeability assay was performed using 1 µM fluorescein (376 kDa, Sigma-Aldrich) for 1 h
Aliquots were taken from the basolateral compartment every 20 min
A parallel assay with cell-free transwells was used to calculate the endothelial permeability coefficient (Pe)

controls

Positive control: Not mentioned
Negative control: Cell-free transwells (used to calculate the endothelial permeability coefficient (Pe))

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