Characterization of arachnoidal cells cultured on three-dimensional nonwoven PET matrix.
OBJECTIVE
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.


CONCLUSIONS
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.
Mehta
Bhavya C
BC
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA.
Holman
David W
DW
Grzybowski
Deborah M
DM
Chalmers
Jeffrey J
JJ
D001099:Arachnoid / Q000166:cytology* / Q000502:physiology*; D040761:Biomimetic Materials / Q000737:chemistry; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000737:chemistry*; D006801:Humans; D010539:Permeability; D011093:Polyethylene Terephthalates / Q000737:chemistry*; D023822:Tissue Engineering / Q000379:methods*
D011093:Polyethylene Terephthalates