Recombinant Spider Silk Membranes Promote Human Renal Epithelial Differentiation and Function

ABSTRACT: Porous membranes are frequently used as supports of cell monolayers in functional studies of epithelial and endothelial barriers. However, conventional polymer-based membranes such as those made of polycarbonate do not mimic the structural and biochemical properties of native basement membranes, which may limit cellular differentiation and function. Here, we use a nanofibrillar membrane made of recombinant spider silk functionalized with the integrin-binding RGD motif of fibronectin and coated with human kidney-specific laminin-521 (LN/FN-silk) as a novel substrate for renal epithelial monolayer cultures. Cellular morphology, mRNA expression, barrier properties and transporter activity were assessed using scanning and transmission electron microscopy, RNA-sequencing, lucifer yellow permeability assays, immunofluorescence staining and fluorescent probe transport assays. Immortalized renal proximal tubular epithelial cell (RPTEC/TERT1) monolayers cultured on LN/FN-silk membranes and in co-culture with human fibroblasts (fHDF/TERT166) exhibited cuboidal morphology, reduced cell death and maintained barrier integrity with tight junction expression compared to conventional polymer-based transwell membranes. In contrast to the LN/FN-silk membranes, the polycarbonate membranes released the endocrine disrupting toxicant bisphenol A, triggering estrogen-mediated signalling. Furthermore, cells on LN/FN-silk membranes showed enhanced directional anion and cation transport, compared to cells cultured on conventional membranes. In summary, the results indicate that LN/FN-silk membranes improve differentiation and functional maintenance of renal epithelial cell cultures, thereby allowing preservation of renal epithelial transport functions.

ORGANISM(S): Homo sapiens

PROVIDER: GSE307014 | GEO | 2026/01/22

REPOSITORIES: GEO