Recreating cellular barriers in human microphysiological systems in-vitro

Abstract

Within cellular barriers, cells are separated by basement membranes (BMs), nanometer-thick extracellular matrix layers. In existing in-vitro cellular-barrier models, cell-to-cell signaling can be preserved by culturing different cells in individual chambers separated by a semipermeable membrane. Their structure does not always replicate the BM thickness nor diffusion through it. Here, a porous polymeric nanofilm made of poly(D-L-lactic acid) (PDLLA) is proposed to recreate the BM in a microfluidic blood-brain-barrier model. Nanofilms showed an average thickness of 275{nm}-25{nm and a maximum pore diameter of 1.6\ μ{m. Human umbilical vein endothelial cells (HUVECs) were cultured on PDLLA. After 7 days, viability was >95% and cell morphology did not show relevant differences with HUVECs grown on control substrates. A protocol for suspending the nanofilm between 2 microfluidic chambers was identified and showed no leakage and good sealing. Clinical Relevance - Preclinical models of cellular barriers are a key step towards a deeper understanding of their roles in pathogenesis of various diseases: a physiologically relevant microfluidic model of the blood brain barrier (BBB) allows high-throughput investigations of BBB contribution in neurodegenerative diseases and cruelty-free screenings of drugs targeting the brain.