Abstract
The blood–brain barrier (BBB) is a selectively permeable barrier that separates the circulating blood from the extracellular fluid of the brain and is an essential component in brain homeostasis. In vitro BBB models are valuable supporting tools that can precede and complement animal and human studies of the development and progression of the central nervous system diseases. At present, mono-, co-, and tri-culture models that use porcine, murine, or human cells have been developed. We have optimized a two-dimensional model of the human BBB using primary human brain microvascular endothelial cells and normal human astrocytes. We have validated the effectiveness of our model with transmigration assays of human blood monocytes toward CCL19, a natural ligand of the chemokine receptor CCR7. This model offers the following advantages:•It is simple, convenient, and requires small quantities of material, reagents, and primary cells.•It can be used to monitor cell migration through the BBB.•It can be used to assess brain capillary permeability in the presence of xenobiotic, pro-inflammatory, or other substances.
Highlights
The blood–brain barrier (BBB) is a selectively permeable barrier that separates the circulating blood from the extracellular fluid of the brain and is an essential component in brain homeostasis
Primary human cells used in co-culture provide a more realistic BBB model than mono-cultures and cell line co-cultures [1,2,3,4]
We used freshly isolated monocytes in chemokine receptor type 7 (CCR7)-dependent transmigration assays to assess the co-culture of human brain microvascular endothelial cells (HBMEC) and normal human astrocytes (NHA) model of cell migration through the BBB
Summary
The blood–brain barrier (BBB) is a selectively permeable barrier that separates the circulating blood from the extracellular fluid of the brain and is an essential component in brain homeostasis. Primary human endothelial cells and astrocytes are co-cultured using ThincertsTM tissue culture inserts to obtain a selective and tight in vitro model of the human BBB. 2. Completely coat 100-mm adherent cell plates with 1.5 or 2.0 mL preheated Attachment Factors for HBMEC and NHA, respectively.
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