title: Topographically-patterned porous membranes in a microfluidic device as an in vitro model of renal reabsorptive barriers.
abstract: Models of reabsorptive barriers require both a means to provide realistic physiologic cues to and quantify transport across a layer of cells forming the barrier. Here we have topographically-patterned porous membranes with several user-defined pattern types. To demonstrate the utility of the patterned membranes, we selected one type of pattern and applied it to a membrane to serve as a cell culture support in a microfluidic model of a renal reabsorptive barrier. The topographic cues in the model resemble physiological cues found in vivo while the porous structure allows quantification of transport across the cell layer. Sub-micron surface topography generated via hot-embossing onto a track-etched polycarbonate membrane, fully replicated topographical features and preserved porous architecture. Pore size and shape were analyzed with SEM and image analysis to determine the effect of hot embossing on pore morphology. The membrane was assembled into a bilayer microfluidic device and a human kidney proximal tubule epithelial cell line (HK-2) and primary renal proximal tubule epithelial cells (RPTEC) were cultured to confluency on the membrane. Immunofluorescent staining of both cell types revealed protein expression indicative of the formation of a reabsorptive barrier responsive to mechanical stimulation: ZO-1 (tight junction), paxillin (focal adhesions) and acetylated α-tubulin (primary cilia). HK-2 and RPTEC aligned in the direction of ridge/groove topography of the membrane in the device, evidence that the device has mechanical control over cell response. This topographically-patterned porous membrane provides an in vitro platform on which to model reabsorptive barriers with meaningful applications for understanding biological transport phenomenon, underlying disease mechanisms, and drug toxicity.
authors: Boston University, Department of Mechanical Engineering, Boston, MA 02215, USA.
authors: Else M
authors: EM
authors: Frohlich
authors: José Luis
authors: JL
authors: Alonso
authors: Jeffrey T
authors: JT
authors: Borenstein
authors: Xin
authors: X
authors: Zhang
authors: M Amin
authors: MA
authors: Arnaout
authors: Joseph L
authors: JL
authors: Charest
mesh_terms: D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism; D008567:Membranes, Artificial; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D051419:Paxillin / Q000378:metabolism; D016062:Porosity; D014404:Tubulin / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D008567:Membranes, Artificial; D051419:Paxillin; D014404:Tubulin; D062826:Zonula Occludens-1 Protein
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