Journal: bioRxiv
Article Title: TNFSF13 insufficiency disrupts human colonic epithelial cell-mediated B cell differentiation
doi: 10.1101/2024.09.23.614260
Figure Lengend Snippet: (A) Schematic of co-culture model. (B) Percentage of plasmablasts differentiated from sorted human memory B cells at day 8 post-seeding via co-culturing with equal numbers of control, VEO-IBD, and variant colonoids in an IntestiCult media and B cell media mixture (ratio 1:1). (C) Percentage of plasmablasts differentiated from sorted human memory B cells at day 8 post-seeding via culturing in conditioned media consisting of B cell media and conditioned media (ratio 1:1). (D-E) Percentage of plasma cells and IgA + plasma cells differentiated from sorted human memory B cells at day 14 post-seeding with B cell media-conditioned media (ratio 1:1). (F) ELISA for IgA in media differentiated from sorted human memory B cells at day 14 post-seeding. (G-H) Percentage of plasma cells and IgA + plasma cells differentiated from sorted human memory B cells at day 14 post-seeding. (I) ELISA for IgA in media from sorted human memory B cells at day 14-post seeding by culturing in conditioned media mixture starting at day 6 post-seeding. P value shown in the bar graphs unless P >0.05. One-way ANOVA (with multiple comparisons) or two-tailed Student’s t -test was used for statistical analysis. n=3 lines of colonoids from 3 different patients for Control and VEO-IBD, n=3 passages/batches of colonoids for Variant. n=3 passages/batches of iPSC-organoids. n=7 independent donors to obtain human memory B cells.
Article Snippet: Colonoids were fed every other day in human IntestiCult Organoid Growth Media (Stem Cell Technologies, British Columbia, Canada) and collected or split roughly 7 days after plating depending on the specific experiment.
Techniques: Co-Culture Assay, Control, Variant Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Two Tailed Test
