Abstract
Purpose: To culture physiologically functional primary arachnoidal cells on a suitable polymer substrate for an in-vitro model of the cerebrospinal fluid outflow pathway. Methods: Primary cultures of arachnoidal cells were prepared within 24 hours post-mortem from brain tissue obtained from human cadavers at autopsy. Arachnoidal cells were characterized using immunocytochemistry and seeded onto needle punched non-woven poly(ethylene terephthalate)(PET) scaffolds. Metabolic rate, cell growth rate in log phase, morphologic assessment, immunocytochemistry, and protein analysis were used to characterize the cultures in both 2-D and 3-D-culture. Functional outflow assessment was performed using the Lucifer Yellow (LY) permeability assay and hydraulic conductivity (Lp) determination. Results: Cells cultured on PET scaffold grew slightly slower than cells grown in 2-D-culture as measured by metabolic rate and growth rate, however, they often formed sheets that bridged between the adjacent scaffold filaments forming many junctional protein connections. LY permeability coefficients of 2-D cells were compared with cells from scaffolds, and were not significantly different (p > 0.05) for both culture conditions. Average Lp of cells from 2-D-culture and 3-D-scaffolds were compared and shown not to be significantly different. Conclusion: Based on the biochemical and functional analysis, it has been shown that cells cultured on 3D-PET scaffolds retained the same properties as cells from 2D-culture plates.
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