Automated production of nerve repair constructs containing endothelial cell tube-like structures

The data that support the findings of this study, entitled Automated production of nerve repair constructs containing endothelial cell tube-like structures

Videos of the automated production of engineered neural tissue (EngNT) constructs using gel aspiration-ejection (GAE) were taken at UCL and McGill University. These data refer to the characterisation of two sizes of EngNT, 12G and 16G, produced using this technique containing human umbilical vein endothelial cells (HUVECs). Tensile testing data was collected from acellular constructs, performed using the Univert Mechanical Testing Instrument with a calibrated 10 N load cell and a controlled displacement rate of 0.1 mm/s. Viability data of engineered neural tissue containing HUVECs (EngNT-HUVEC) covers local viability, using LIVE/DEAD^TM Cell Imaging Kit (488/570), and whole construct viability, using Lactase Dehydrogenase (LDH) Assay Kit (Cytotoxicity) collected using a NanoZoomer S60 Digital Slide Scanner and a SpectraMAX M5 Multi-Mode Microplate Reader, respectively. CellTiter-Glo® 3D Cell Viability Assay data reports the ATP concentration in EngNT-HUVEC constructs relative to their starting hydrogels, measured using a SpectraMAX M5 Multi-Mode Microplate Reader. Alignment of cells within EngNT-HUVEC was captured by Volocity^TM v6.5.1 analysis of 20 µm z-stacks taken using Zeiss LSM 710 confocal microscope at predetermined positions along the construct stained with Rhodamine 110 Phalloidin and Hoechst 33342. Viability, metabolic activity and alignment assays were performed 24 h after EngNT-HUVEC construct formation. Neurite length and alignment data were collected using ImageJ and Volocity^TM v6.5.1, respectively, analysing the co-culture of neurons on the EngNT-HUVEC and acellular constructs imaged using Zeiss LSM 710 confocal microscope of samples stained with anti-betaIII tubulin and Hoechst 33342. EngNT-HUVEC made from starting hydrogels with two seeding densities (0.5 mill/mL and 2.0 mill/mL) were cultured for 2 or 4 days. These samples were stained with Rhodamine 110 Phalloidin and Hoechst 33342 and analysed for inner angiogenic features, using ImageJ plugin Angiogenesis Analyzer, and outer endothelialisation, using Volocity^TM v6.5.1. 16G EngNT-HUVEC constructs were further analysed for lumen number from 10 µm sections stained with Rhodamine 110 Phalloidin and Hoechst 33342, imaged on Zeiss AxioLab A1 fluorescent microscope. RT-PCR data from EngNT-HUVEC were collected using GoScript Reverse Transcriptase Kit (Promega) and Power SYBRä Green PCR Master Mix using SimpliAmp Thermal Cycler and QuantStudio3 instrument, analysed by QuantStudio Design and Analysis Software v1.5.1 with hypoxanthine guanine phosphoribosyl transferase (HPRT1), ribosomal protein lateral stalk subunit P0 (RPLP0) and ribosomal protein S18 (RPS18) reference genes.

Data collected between December 2022 and January 2024.