Abstract 495: Tissue Engineering Of A Three-layered Artery

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A human-induced pluripotent stem cell (hiPSC)-3D artery mimicking the multi-layered anatomy of blood vessel has not yet been established. Here, we aimed to engineer arteries consisting of human endothelial cells (EC), vascular smooth muscle cells (VSMC), and fibroblasts, correspondingly mimicking the main cellular components of the intima, media, and adventitia of a blood vessel. We differentiated hiPSC to obtain EC, VSMC, and fibroblasts. A 3D printing-based molding technique was used to engineer the circular structure of an artery. For the media-like layer, hiPSC-VSMCs mixed with collagen were seeded into 3D-printed polylactic acid molds with a centered needle serving as supporting structure. The adventitia-like layer was engineered around the VSMC-layer using the same method but with fibroblast-collagen mixture. After formation of the fibroblast layer, the needle was removed and hiPSC-ECs were coated on the VSMC lumen by rotational cell injection. Brightfield, fluorescence and two-photon microscopy were utilized to visualize cell arrangement, EC coverage, and collagen usage and organization. RNA sequencing (RNAseq) was harnessed to molecular profile cells in 2D (Petri dish) and 3D culture (engineered arteries). With 10M hiPSC-VSMCs, 10M hiPSC-fibroblasts, and 0.2M hiPSC-EC, we successfully engineered the three-layered “artery”. The built-up took 16 days. The condensation of cell-collagen mixture likely relied on the rearrangement of collagen fibers by the cells through enzymic reactions in the extracellular matrix. Immunostaining of alpha-SMC actin indicated rod-shaped hiPSC-VSMC aligned in the artificial tubes. Two-photon microscopy revealed over 95% coverage of hiPSC-EC of the luminal wall. Transcriptome data suggested that 3D culture promotes the expression of genes of the calcium signaling pathway and leads to downregulation of genes of the atherosclerosis pathway in SMC. Taken together, the human-induced pluripotent stem cell (hiPSC)-3D artery might serve as a model for functional studies and experimental models of vascular injury and inflammation. In addition, our three-layered tube - mimicking a human artery - seems to improve the molecular profile of vascular cells derived from hiPSC as compared to 2D culture.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.