Fig. 3: Consideration for modeling the ST barrier.
From: Trophoblast stem cell-based organoid models of the human placental barrier
A A schematic diagram of fabrication of a column insert and cell seeding. B An image of the column insert. C Schematic of ST barrier formation. D Culture schedules for ST barrier models. Triangular marks indicate medium exchange. E Transepithelial/transendothelial electrical resistance (TEER) values for the barrier models. F–J Immunofluorescence staining images of SDC1 and E-cadherin. F Percentage of the area covered with SDC1-expressing cells in the circular collagen membrane in each sample was determined using ImageJ version 1.47t (National Institutes of Health) (N = 1). G A partially magnified image of Fig. 3F(a). TS cells on the column device were cultured in PreM for 2 days, W-DM for 2 days, and S-DM for 3 days. H A 3D structure and (I) cross-sections in ST barrier models analyzed using a Zeiss LSM 700 confocal microscope (Carl Zeiss). A sample that is shown in Fig. 3F(a) was analyzed. J Immunostaining of a frozen section of the ST barrier models. K TEER levels for the ST or TS models. TS cells were seeded at 1.2 ×106 cells/mL ×40 μL/column on day 0 in this experiment (N = 7, STM; N = 3, TSM). *P < 0.05, Student’s t test (STM vs. TSM). STM, The three kinds of medium shown in Fig. 3D(a). TSM, trophoblast stem cell medium. Data are shown as mean ± SE. L Levels of secreted hCG (N = 3). *P < 0.05, Tukey’s multiple comparison test. Data are shown as mean ± SE. The scale bars indicate 1 cm (B), 200 μm (F), or 100 μm (I and J). F, G, J Images were taken using BZ-X800/810. N refers to replicates (biologically independent samples).
