Generation of a Neurovascular Unit

Take neural stem cells or NSCs and brain microvascular endothelial cells or BMECs in the desired ratio.

Suspend the cell mixture in an extracellular matrix or ECM solution.

Spread the mixture on the bottom of a confocal dish and incubate.

The ECM polymerizes, forming a three-dimensional scaffold that enables cellular adhesion.

Spread a layer of NSCs in the ECM solution over the solidified layer.

Incubate to polymerize ECM. This mimics the layered brain tissue, enhancing cell-cell interactions.

Add media and incubate.

The growth factors in the media promote cell proliferation while facilitating BMECs to organize into vascular-like networks, forming a neurovascular unit or NVU.

Replace the media with media containing vitamin A, facilitating differentiation of NSCs into neurons and glial cells.

The cells within NVU are spontaneously assembled into a modular organization replicating a brain-like microenvironment.