Human Brain Microvascular Endothelial Cells and Umbilical Vein Endothelial Cells Differentially Facilitate Leukocyte Recruitment and Utilize Chemokines for T Cell Migration

Figure 3

THBMEC cultures are more restrictive towards PBMCs transmigration than HUVECs. 10⁶ of PBMCs were added into the transwell inserts containing confluent THBMECs or HUVECs and allowed to transmigrate for the indicated time. Migrated cells were collected and three color stainings were performed in one step using anti-CD3 PerCP and anti-CD14 APC antibodies. Data were collected with an LSR flow cytometer and analyzed using FlowJo® software. Parallel migration assays were performed using calcein-AM labeled and unlabeled PBMCs, and migration ratios calculated as stated in Section 2. The assays were performed in quadruplicate using four different donors. Confluent HUVEC cultures are more permissive towards PBMCs migration than THBMECs. ^*indicates 𝑃<.05 comparing THBMECs with HUVECs. PBMCs transmigration across these endothelial cultures follows the pattern: monocytes > T cells > B cells (data about B cells not shown).