Development of a living membrane comprising a functional human renal proximal tubule cell monolayer on polyethersulfone polymeric membrane

• Published in: Acta biomaterialia Vol. 14; pp. 22 - 32
• Main Authors: Schophuizen, Carolien M.S., De Napoli, Ilaria E., Jansen, Jitske, Teixeira, Sandra, Wilmer, Martijn J., Hoenderop, Joost G.J., Van den Heuvel, Lambert P.W., Masereeuw, Rosalinde, Stamatialis, Dimitrios
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.03.2015
• Subjects: Cations; Cell Adhesion - drug effects; Cell Line, Transformed; Coated Materials, Biocompatible - pharmacology; Coating; Collagen Type IV - pharmacology; Creatinine; Devices; Human; Human kidney proximal tubule cells; Humans; Immunohistochemistry; Kidney Tubules, Proximal - cytology; Kidneys; l-DOPA; Levodopa - pharmacology; Living membrane; Membrane biofunctionalization; Membranes; Membranes, Artificial; Microscopy, Electron, Scanning; Monolayers; Permeability; Polyethersulfones; Polymers - pharmacology; Renal assist device; Sulfones - pharmacology; Water; l-DOPA; Renal assist device; Living membrane; Human kidney proximal tubule cells; Membrane biofunctionalization
• ISSN: 1742-7061; 1878-7568
• DOI: 10.1016/j.actbio.2014.12.002

[Display omitted] The need for improved renal replacement therapies has stimulated innovative research for the development of a cell-based renal assist device. A key requirement for such a device is the formation of a “living membrane”, consisting of a tight kidney cell monolayer with preserved functional organic ion transporters on a suitable artificial membrane surface. In this work, we applied a unique conditionally immortalized proximal tubule epithelial cell (ciPTEC) line with an optimized coating strategy on polyethersulfone (PES) membranes to develop a living membrane with a functional proximal tubule epithelial cell layer. PES membranes were coated with combinations of 3,4-dihydroxy-l-phenylalanine and human collagen IV (Coll IV). The optimal coating time and concentrations were determined to achieve retention of vital blood components while preserving high water transport and optimal ciPTEC adhesion. The ciPTEC monolayers obtained were examined through immunocytochemistry to detect zona occludens 1 tight junction proteins. Reproducible monolayers were formed when using a combination of 2mgml−1 3,4-dihydroxy-l-phenylalanine (4min coating, 1h dissolution) and 25μgml−1 Coll IV (4min coating). The successful transport of 14C-creatinine through the developed living membrane system was used as an indication for organic cation transporter functionality. The addition of metformin or cimetidine significantly reduced the creatinine transepithelial flux, indicating active creatinine uptake in ciPTECs, most likely mediated by the organic cation transporter, OCT2 (SLC22A2). In conclusion, this study shows the successful development of a living membrane consisting of a reproducible ciPTEC monolayer on PES membranes, an important step towards the development of a bioartificial kidney.