Application of combined gene and cell therapy within an implantable therapeutic device for the treatment of severe haemophilia A [Abstract]

Application of combined gene and cell therapy within an implantable therapeutic device for the treatment of severe haemophilia A [Abstract

HemAcure: Application of combined gene and cell therapy within an implantable therapeutic device for the treatment of severe haemophilia A

HemAcure: Application of combined gene and cell therapy within an implantable therapeutic device for the treatment of severe haemophilia

Biochemical analysis of LO culture medium.

<p>Albumin (A), urea (B) and glucose (C) production at different time points for liver organoids on different substrates. Significant differences at the same time points (<i>p</i><0.05) are marked with an asterisk.</p

Integrin-ß1 (green, 1^st column) and Connexin-32 (red, 2^nd column) expression on Matrigel-LSEC, Matrigel-PBS, ECM-LSEC and ECM gel substrates.

<p>Blue: cell nuclei (DAPI). Third column: merged image channels. Scale bar: 10 μm.</p

Modelling LO oxygen transport and consumption.

<p>A) Results from the computational model, using the measured OCR_LO at 24 h. LO in the static well (left) and in the LiveBox1 (right) The computed velocity field in the bioreactor is also shown. B) Oxygen concentration along the vertical axis passing through the centre of the LO (z-axis) cultured either in the LiveBox1 bioreactor (solid line) or in the static well (dashed line). The dotted horizontal line indicates the threshold for hepatocyte function (i.e. 0.02 mM).</p

Measurement and analysis of LO oxygen consumption.

<p>A) Time-dependent oxygen profile at 24 h after cell seeding. B) Oxygen concentration versus time data measured in the first 15 minutes used to derive the <i>OCR</i>_<i>LO</i> via a linear fit (dashed line).</p

Boundary conditions used for the oxygen convection and diffusion and for the Navier-Stokes models.

<p>Boundary conditions used for the oxygen convection and diffusion and for the Navier-Stokes models.</p

Agarose and liver ECM gels mechanical properties.

<p>A) Compressive moduli of agarose gels prepared at different weight/volume (w/v) concentrations. Agarose gels were not formed below 0.1% w/v. The dashed line represents the stiffness of Matrigel-x2 substrates (estimated from the literature). B) Mechanical properties of 20 mg/mL ECM gel and 10 mg/mL ECM-LSEC gel. The two dashed lines represent the stiffness of 0.1% agarose and Matrigel x2 substrates.</p

Mean Pixel Intensity (MPI) for green (Integrin-β1) and red (Connexin-32) channels.

<p>The MPI was calculated from n = 3 independent experiments per substrate investigated, imaging and analysing n = 3 different DAPI-positive areas in each sample (randomly selected). Different letters indicate significant differences between groups (one-way ANOVA, <i>p</i> < 0.05): lowercase letters are referred to ANOVA results of green MPI, while capital letters denote ANOVA results of red MPI. Note that the green and red MPI levels cannot be compared due to differences in the laser power used and in the fluorophore excitation/emission efficiencies.</p

Outcome of liver organoid formation on different substrates at 24 h.

<p>Outcome of liver organoid formation on different substrates at 24 h.</p