Journal: bioRxiv

Article Title: Efficient Generation of Functional TCRαβ + Cytotoxic T Cells from hiPSCs via Small-Molecule Modulation

doi: 10.64898/2026.03.31.715684

Figure Lengend Snippet: A: Schematic overview of the screening approach. hiPSC-derived CD34 + hemogenic endothelial cells were differentiated towards ProT cells for 14 days in a stroma-free protocol . At iT day 14 cells were harvested and replated into a 384 well plate and exposed to a small molecule library. After 7 days cells were stained with anti CD56, anti CD4, anti CD7 with 7-AAD and Calcein Violet viability dyes. Scale bar indicates 20 µm. B: Primary screening results. 5 hits were identified when a Z-score threshold of 1 was applied. C: Hit validation, bar graphs depicting mean percentage of CD4 + cells out of CD45 + population on iT day 28 in hiPSC line EZH1 mut/wt (screening line), BCH1157 and BCH1566. Error bars depict SEM of replicate wells, one way ANOVA comparison against DMSO. E: Exemplary flow cytometry plots on iT day 28 demonstrating an increased proportion of CD4 + CD8 + DP cells after GNF351 treatment in BCH1157 hiPSC-derived ProT cells. F: Mean values plotted from two independent experiments with either continuous drug treatment (solid circles) or one time drug exposure (open circles). SEM of technical replicates is indicated by blue shading. Two-way ANOVA with Dunnett’s multiple comparison test against respective untreated cells.

Article Snippet: The cells were then filtered through a filter-top polystyrene flow cytometry tube (Fisher Scientific, #08-771-23), washed with flow cytometry buffer (PBS + 2% FBS + antibiotics), and centrifuged at 300xg for 5 minutes followed by antibody staining and flow cytometric analysis for expression of CD45 and CD19.

Techniques: Derivative Assay, Staining, Biomarker Discovery, Comparison, Flow Cytometry

Journal: STAR Protocols

Article Title: Protocol to annotate dendritic cell maturation types in vivo making use of lipid nanoparticle-based approaches

doi: 10.1016/j.xpro.2026.104395

Figure Lengend Snippet: Panel design For efficiency, staining mixes are prepared one day before the readout. These include mixes for experimental samples, single-stained cell controls, and single-stained bead controls ( C and 2D). Mixes are prepared without cells and added on the day of the readout. The optimized panel uses two sequential staining steps. (A) 96-well plate layout for single-stained beads (B). (B) 96-well plate layout for the staining for the single-stained cell controls, SS1: single-stained staining 1, SS2: singe-stained staining 2. (C) Panel overview and information on staining mixes for the experimental samples, single-stained beads and single-stained cell controls for conventional flow cytometry. (D) Panel overview and information on staining mixes for the experimental samples, single-stained beads and single-stained cell controls for FACS. We assign a number to each marker to simplify labeling of the wells for the single-stained controls. The total volume of the experimental staining mixes for experimental samples is determined by the number of samples to be stained. For single-stained controls, we always prepare the staining in a total volume of 200 μL. All staining mixes and control samples are then prepared according to the required antibody dilutions. For the channel corresponding to the LNPs, single-stained controls consist of cells from a mouse injected with LNPs. For single-stained beads, an antibody conjugated to a fluorochrome detected in the same channel need to be used.

Article Snippet: Add 500 μL of the 2× digestion buffer 1. c. Incubate the Eppendorf tube in a 37°C heat block and put the shaker on maximum for 20 min. d. Resuspend tissue every 10 min. e. Filter over a 5 mL polystyrene FACS tube with filtercap (35 μM), smash, wash Eppendorf tube with 2 mL 1× PBS and filter again over the filtercap (35 μM). f. If enrichment for cDCs is necessary, go further to step 11. g. Remove supernatant and resuspend in 200 μL FACS buffer. h. Count cells.

Techniques: Staining, Flow Cytometry, Marker, Labeling, Control, Injection