collagen matrices  (Thermo Fisher)

Journal: Advanced Science

Article Title: Engineering Adaptive Immunity in 3D: A Patient‐Specific Lymphoid Model Using Stromal Networks and Peripheral Blood Mononuclear Cells

doi: 10.1002/advs.202513245

Figure Lengend Snippet: Differentiation of Adipose Derived Stem Cells (ADSC) into Fibroblastic Reticular‐like Cells (FRC). A) Schematic of approaches, P1 & P2, used to differentiate ADSCs into FRC‐like cells. The 10‐day differentiation was followed by analysis using imaging, flow cytometry, and RNA‐sequencing. Created in https://BioRender.com B) i) Representative images showing full well images of ADSCs, FRC P1 , and FRC P2 cells after 10 days of culture within collagen matrices. Scale bar: 5 mm ii) Images taken at x4 on Lionheart microscope stained with Phalloidin 594 (red) and Hoeschst (blue) showing the differences in morphology of the three cell types. Scale bar: 500 µm iii) Images taken at x10 on LEICA STED stained with Phalloidin 594 (red) and Hoeschst (blue) of FRC P1 and FRC P2 . Scale bar: 500 µm. A 3D rotational video of the FRC P1 edge appears as a video in the . C) i) Flow cytometry derived UMAP plot of populations in ADSCs, FRC P1 , and FRC P2 cells after 10 days of differentiation. Each plot represents 16 concatenated samples from four distinct ADSC donors. Values below each UMAP represent the percentage of each subpopulation within the treatment condition. Colors correspond to the subpopulation in the heatmaps. ii) Heat map showing the expression levels of CD21, ICAM1, PDPN, Thy1, CD44, LTBR, and VCAM1 in each population. Values are normalized within each column relative to the specific marker. D) Representative images taken at 20x on Olympus IX83 with immunofluorescence staining of ADSCs, FRC P1 , and FRC P2 for nuclear marker Hoechst (cyan), PDPN (red), and ICAM1 (green). Overlays demonstrate co‐localization of PDPN and ICAM1 within multicellular clusters.

Article Snippet: The 3D collagen matrices were then washed three times with phosphate buffer saline (PBS; Thermo Fisher Scientific Inc, Leicestershire, UK) and placed into 24‐well plates (Thermo Fisher Scientific Inc, Dreieich, Germany).

Techniques: Derivative Assay, Imaging, Flow Cytometry, RNA Sequencing, Microscopy, Staining, Expressing, Marker, Immunofluorescence

Journal: Advanced Science

Article Title: Engineering Adaptive Immunity in 3D: A Patient‐Specific Lymphoid Model Using Stromal Networks and Peripheral Blood Mononuclear Cells

doi: 10.1002/advs.202513245

Figure Lengend Snippet: Embedded Fibroblastic Reticular‐like Cells (FRC) Remodel 3D Matrices. A) i) Representative images, taken every 24 h, showing full well images of ADSCs, FRC P1 , and FRC P2 cells within collagen matrices during the 10‐day differentiation period. Images begin at 60 h when the first notable changes appear. Scale bar: 5 mm ii) Quantified change in matrix area over time for 10 days iii) Quantification of matrix area after 10 days of differentiation. B) Heatmap of extracellular matrix related genes clustered into two groups using k‐means clustering. C) Upregulated KEGG, GO Biological Processes, GO Molecular Function, and GO Cellular Component pathways of enriched genes in i) group 1 (ADSC vs FRC) and ii) group 2 (FRC vs ADSC) of the k‐means clustering heatmap.

Article Snippet: The 3D collagen matrices were then washed three times with phosphate buffer saline (PBS; Thermo Fisher Scientific Inc, Leicestershire, UK) and placed into 24‐well plates (Thermo Fisher Scientific Inc, Dreieich, Germany).

Techniques: