Journal: bioRxiv

Article Title: Early-life mucosal T cells direct intestinal stem cell fate via a coordinated developmental program

doi: 10.64898/2026.05.08.723752

Figure Lengend Snippet: (A) Co-culture strategy (created with Biorender.com ). Small intestine (SI) was cryopreserved, and stable organoid lines were generated. T cells were isolated from SI tissue (B-E, H-K) or cord blood (F-G) by positive selection. CD3+ cells were co-cultured with established SI organoid lines generated from fetal (B-I) , neonatal (D-E) , or adult (I-K) donors. (B, D, F, H) Representative images from day 6 of co-culture. Scale bar is 530 µm. (C) Data were generated from one organoid and T cell donor per fetal age group and analyzed by one-way ANOVA with Tukey’s post-hoc test. Only significant comparisons are denoted. Data are shown as means ± SEM. (E, G, I) Data were generated from 2-4 organoid and T cell donors each. Data were analyzed via Mann-Whitney and are shown as medians ± IQR. (K) Data were generated from 3 organoid donors and 4 T cell donors and are normalized to the number of organoids generated in mock (no T cell) conditions, represented as fold change. Each dot represents the average of three technical replicate wells (C) or one well of organoids (E, G, I, K) . Data were analyzed via Welch’s t-test and are shown as means ± SEM. Each experiment was repeated 2-3 independent times. Each dot represents one well of organoids. Significance is indicated as follows: * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001; ns, not significant (p>0.05).

Article Snippet: T cells were isolated via CD3 microbead positive selection (Miltenyi, cat#130-050-101 or 130-097-043) via MS or LS columns (Miltenyi, cat#130-042-201 or 130-042-401) per manufacturer’s instructions (Miltenyi).

Techniques: Co-Culture Assay, Generated, Isolation, Selection, Cell Culture, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Proinflammatory Cytokine Preconditioning Enhances the Therapeutic Potency of Different Types of MSCs in Inflammation

doi: 10.3390/ijms27094090

Figure Lengend Snippet: The effects of naïve and Cytomix-stimulated MSCs on T cell proliferation. The gating strategy is shown in . ( A – C ) BM-MSCs, iMSCs WT and iMSCs B2M KO inhibition on the expansion of CD3+, CD3+CD4+ and CD3+CD8+ T cells via CM. Cytomix enhanced the three types of MSCs’ inhibition on the EI of T cells. ( D – F ) All types of MSCs significantly inhibited the division of CD3+, CD3+CD4+ and CD3+CD8+ T cells when co-cultured with PBMCs, in a dose-dependent manner. ( G – I ) All of the MSCs, naïve or Cytomix stimulated, significantly reduced the EI of CD3+, CD3+CD4+ or CD3+CD8+ T cells, at the lowest ratio of MSCs, with iMSCs WT both naïve and Cytomix-stimulated presenting the strongest inhibition. ( J ) Co-culturing all types of MSCs, naïve or Cytomix-stimulated, increased the production of G-CSF in the media, with a dose-dependent effect. ( K ) All types of MSCs significantly decreased the level of IFN-γ at the ratios of 1:20 and 1:10, while the three types of naïve MSCs significantly decreased it at 1:50. ( L ) Naïve and Cytomix-stimulated iMSC WTs significantly increased the level of TNF-α at the ratios of 1:50, 1:20 and 1:10, while Cytomix-stimulated iMSC B2M KO significantly increased it at the ratio of 1:50 and 1:20. ( M ) Naïve BM-MSCs significantly increased the level of IL-10 production at the ratios of 1:20 and 1:10. Representative results of four independent experiments are shown. Results are shown as mean ± SD ( n = 3 in CM; n = 4 in co-culture, each group), and analysed by two-way ANOVA with Šídák’s multiple comparisons test to compare naïve vs. Cytomix in each type of MSCs (no significance in all of the settings), or Tukey’s multiple comparisons test to compare different ratios of MSCs to PBMCs in each MSC group, or Dunnett’s multiple comparisons test to compare different MSC groups vs. vehicle in the setting of the same ratio. */**/***/**** represent comparisons of each type of MSC vs. vehicle; #/## represent comparisons of different ratios of each cell type with different levels of p < 0.05/0.01. EI: expansion index; DP: division percentage.

Article Snippet: The PBMCs were stained with anti-human CD3-APC-Vio770, CD4-APC and CD8-PE-Vio770 antibodies (Cat# 130-113-136, 130-113-222, 130-110-680; Miltenyi Biotec) for 30 min on ice in the dark.

Techniques: Inhibition, Cell Culture, Co-Culture Assay