Biomedical Engineering Reference
In-Depth Information
a
b
c
d
Fig. 11 Brain endothelial cells (bEND3) expressing GPR124 were induced to form sprouts by
gradients of forebrain-conditioned medium. a Schematic of microfluidic device to induce sprouting
morphogenesis within a gradient of conditioned medium (CM). b A representative sprout cultured
within a gradient of CM from E12.5 forebrain cortical neurons. c, d Directional sprouting
morphogenesis is enhanced in cultures expressing GPR124 (+) and exposed to gradients of
forebrain-conditioned medium (F). Cells treated with GPR124 shRNA (sh) or exposed to gradients
of hindbrain-conditioned medium (H) resulted in significantly less sprouting morphogenesis and
did not display pathfinding behavior ( p \ 0.05). Originally published in [
72
]
modified to enable both 2D and 3D endothelial cell culture (Fig.
11
a). In this
assay, bEnd3 cells were initially cultured as a monolayer within a side chamber that
was connected to the main culture chamber though 30 lm micro-capillaries. The
main culture chamber was filled with a 3D matrix blend of collagen I (1.9 mg/mL)
with fibronectin (50 lg/mL). The cells undergo 2D migration through the micro-
capillaries followed by sprouting morphogenesis into the 3D matrix, where they are
exposed to a stable gradient of paracrine secretions from cultured cortical cells of
murine forebrain origin (Fig.
11
b). This gradient is oriented perpendicular to the
initial sprouting direction. Total sprout length was specifically increased by
Search WWH ::
Custom Search