Fig. 4: Rapid neural differentiation in perfused systems.
From: Large-scale perfused tissues via synthetic 3D soft microfluidics
a Representative experimental results demonstrating the result of 2 days of neuronal induction in control organoids (left column), non-perfused (middle column) and perfused (right column) tissue constructs. Top, middle and bottom row: immunofluorescent images of stem cell marker NANOG, early neural marker PAX6 and cell adhesion proteins N-Cadherin (green) and E-Cadherin (red). The images represent transverse cross-sections of the tissue constructs. In most cases, the cross-sections of capillaries are visible as circular structures. Statistics on biological replicates are given in b. b Average expression of PAX6 (Ctrl: Not detected, n = 5; NP: 0.2%±0.2%, pControl = 0.35(n.s.), n = 9; P: 21.5 ± 2%, pControl = 0.00002, n = 12), NANOG (Ctrl: 73.6 ± 8%, n = 9; NP: 16.1 ± 2%, pControl = 0.00007, n = 11; P: not detected, n = 10), NCad (CDH2) (Ctrl: 100 ± 1%, n = 8; NP: 7.6 ± 1%, pControl = 4e-13, n = 3; P: 55.7 ± 9%, pControl = 0.009, n = 5) and ECad (CDH1) (Ctrl: 99 ± 2%, n = 12; NP: 26.6 ± 7%, pControl = 0.0001, n = 6; P: 4.5 ± 2%, pControl = 2e-10, n = 5) markers in control organoids (Ctrl), non-perfused (NP) and perfused (P) constructs. Data are derived from independent experiments and are represented as mean ± SEM, asterisks (*) denote statistical significance between control organoids and tissue constructs (unpaired two-tailed Student’s t-test, 95% confidence interval). c UMAP plot of the combined dataset showing the localization of cells from control organoids, non-perfused and perfused constructs in the UMAP space. Color bars represent scaled level of expression of the corresponding genes. d UMAP plot of the combined dataset highlighting locations of PAX6, NANOG, NCad (CDH2) and ECad (CDH1) expressing cells in the UMAP space. Scale bar: 250 µm.
