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Deborah M. Grzybowski, David W. Holman, Steven E. Katz, Martin Lubow; In Vitro Model of Cerebrospinal Fluid Outflow through Human Arachnoid Granulations. Invest. Ophthalmol. Vis. Sci. 2006;47(8):3664-3672. doi: 10.1167/iovs.05-0929.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To describe and validate an in vitro model of the arachnoid granulation (AG) outflow pathway for cerebrospinal fluid (CSF), by using human AG cells grown on a filter membrane support and perfused in a modified Ussing chamber at pressures analogous to normal human intracranial pressures.
methods. Human AG cells were grown, characterized, seeded onto filter membranes, and perfused in the physiologic (basal to apical, B→A) or nonphysiologic (apical to basal, A→B) directions. Cells were fixed under pressure after perfusion and prepared for electron microscopy (EM).
results. The average cellular hydraulic conductivity in the B→A direction (10 total) was 4.52 ± 0.43 μL/min per mm Hg/cm2 with an average transcellular pressure decrease of 3.13 ± 0.09 mm Hg. The average cellular hydraulic conductivity in the A→B direction (six total) was 0.29 ± 0.16 μL/min per mm Hg/cm2 with an average transcellular decrease in pressure of 3.33 ± 0.16 mm Hg. Cells perfused nonphysiologically showed a large number of dead and dying cells. EM postperfusion analysis showed that AG cells were integrally attached to the underlying filter membrane. Large extracellular cisternal spaces were visible between overlapping AG cells and vacuoles within the cytoplasm. It is possible that these spaces within and between cells represent pathways for transcellular and paracellular transport of fluid.
conclusions. The results demonstrate that AG cells in vitro show a statistically significant greater flow rate and cellular hydraulic conductivity when perfused in the physiologic versus the nonphysiologic direction under normal intracranial pressures. These results suggest that this in vitro model of the AGs can accurately replicate the unidirectional flow of CSF in vivo.
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