Towards 3D Bioprinting of a Vascularized Convoluted Proximal Tubule
Abstract
Three-dimensional (3D) constructs that recapitulate native tissue architectures offer biomimetic microenvironments in which to perform improved pharmacological studies of drug efficacy and toxicity. The convoluted proximal tubule (PT) is the principal site implicated in renal disease progression and presents an ideal target for disease modeling and therapeutics. Although previous investigators have developed 3D structures that reproduce specific proximal tubule morphologies and functions, these models have been limited by their lack of integrated vasculature to simulate crucial reabsorption and transport processes between the proximal tubule and adjoining blood vessels. This thesis sought to expand the functionality of current proximal-tubule-on-chip technologies by 3D bioprinting a vascularized PT. The model explored in this thesis incorporated cellular compatibilities, extracellular matrix formulations, and geometric design considerations to demonstrate co-culture of adjacent endothelialized and epithelialized tubules for three weeks under 3D-perfused flow. The MTS cell proliferation assay was utilized to select glomerular microvascular endothelial cells expressing red fluorescent protein (RFP-GMECs) as the vascular cell line for 3D perfused co-culture with immortalized proximal tubule epithelial cells (PTECs-Terts). The RFP-GMECs and PTEC-Terts expressed appropriate vascular and renal cell markers when cultured on extracellular matrices statically and under flow. Transepithelial transport of locked-nucleic acid across the proximal tubule channel lining was further demonstrated in a non-endothelialized model. Ultimately, RFP-GMECs and PTEC-Terts were successfully co-cultured in a vascularized PT model for three weeks. The vascularized PT construct presented here enables functional assays of active transport processes, analysis of nephrotoxic drug effects, and exploration of altered tubular architectures. Various biological and technical considerations can be individually addressed through this customizable 3D model, permitting analysis of engineering principles essential to developing more complicated versions of the vascularized proximal tubule and, ultimately, larger scale renal tissue.Terms of Use
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http://nrs.harvard.edu/urn-3:HUL.InstRepos:38811514
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