Journal: bioRxiv

Article Title: A novel approach for reliable differentiation of lymphatic endothelial progenitor cells in vitro

doi: 10.1101/2025.09.23.677051

Figure Lengend Snippet: Stromal vascular fraction (SVF)-derived lymphatic endothelial progenitor cells (LEPCs) show better differentiation by flow cytometry analysis than bone marrow (BM)-derived LEPCs. (A) Flow cytometry workflow of the SVF-derived LEPCs from the first population until the PDPN-positive population. (A.I) PDPN expression histograms showing the expression level for SVF-LEPCs and -MSCs. (A.II) Median fluorescence intensity (MFI) of PDPN in SVF-LEPCs and –MSCs (mean ± SD). (B) Flow cytometry workflow of the BM-derived LEPCs from the first population until the PDPN-positive population (B.I) PDPN expression histograms showing the expression level for BM-LEPCs and -MSCs. (B.II) MFI of PDPN in BM-LEPCs and –MSCs (mean ± SD). Statistical significance: * p -value<0.5, ** p -value<0.01. Mann-Whitney test. n=6 for SVF-LEPCs and n=5 for BM-LEPCs. Mean ± SD represented.

Article Snippet: Human dermal lymphatic endothelial cells (HDLEC, C-14021, PromoCell) were cultured according to the providers’ guidelines.

Techniques: Derivative Assay, Flow Cytometry, Expressing, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: A novel approach for reliable differentiation of lymphatic endothelial progenitor cells in vitro

doi: 10.1101/2025.09.23.677051

Figure Lengend Snippet: Characterization of lymphatic endothelial progenitor cells (LEPCs) shows correct differentiation by overexpression of lymphatic lineage specific markers. (A) RT-qPCR shows statistically significant overexpression of key lymphatic markers such as Pdpn , Vegfr3 , Lyve1 , Vegf-c and Igf2 . n=4. MSC, mesenchymal stem cells. Mean ± SD. (B) Median fluorescence intensity analysis of immunofluorescence staining images shows statistically significant differential expression of PDPN between MSCs and LEPCs. Scale bar: 100 µm. n=6. Statistical significance: * p -value<0.05; ** p -value<0.01. Mann-Whitney test.

Article Snippet: Human dermal lymphatic endothelial cells (HDLEC, C-14021, PromoCell) were cultured according to the providers’ guidelines.

Techniques: Over Expression, Quantitative RT-PCR, Fluorescence, Immunofluorescence, Staining, Quantitative Proteomics, MANN-WHITNEY

Journal: bioRxiv

Article Title: A novel approach for reliable differentiation of lymphatic endothelial progenitor cells in vitro

doi: 10.1101/2025.09.23.677051

Figure Lengend Snippet: Stromal vascular fraction (SVF)-derived lymphatic endothelial progenitor cells (LEPCs) show a reliable lymphatic phenotype in vitro. (A) Genotype stability results of the key overexpressed genes over passages (P), assessed by RT-qPCR. n=3. (B) Scratch-wound assay of the SVF-derived LEPCs in comparison to human dermal lymphatic endothelial cells (HDLECs) along two days in culture. n=3. Scale bar: 200 µm. Mean ± SD. Mann-Whitney test.

Article Snippet: Human dermal lymphatic endothelial cells (HDLEC, C-14021, PromoCell) were cultured according to the providers’ guidelines.

Techniques: Derivative Assay, In Vitro, Quantitative RT-PCR, Scratch Wound Assay Assay, Comparison, MANN-WHITNEY

Journal: bioRxiv

Article Title: A novel approach for reliable differentiation of lymphatic endothelial progenitor cells in vitro

doi: 10.1101/2025.09.23.677051

Figure Lengend Snippet: Stromal vascular fraction (SVF)-derived lymphatic endothelial progenitor cells (LEPCs) show better in vitro angiogenesis phenotype than human dermal lymphatic endothelial cells (HDLEC). (A) LIVE/DEAD staining images of SVF-derived LEPCs and HDLECs culture on a Matrigel (3 mg/mL) coating for 5 hours (2D angiogenesis assay). (B) Angiogenesis analysis of SVF-derived LEPCs compared to HDLECs in a 2D environment. N=5. (C) Bright field images of SVF-derived LEPCs and HDLECs embedded in a 3D Matrigel hydrogel (3 mg/mL) and cultured for 10 days. (D) LIVE/DEAD staining images of SVF-derived LEPCs and HDLECs embedded in a 3D Matrigel hydrogel (3 mg/mL) and cultured for 10 days. (E) Angiogenesis analysis of SVF-derived LEPCs compared to HDLECs in a 3D environment. n=4. Scale bars: (A) 500 µm; (C-D) 200 µm. Statistical significance: * p -value<0.05. Mann-Whitney test.

Article Snippet: Human dermal lymphatic endothelial cells (HDLEC, C-14021, PromoCell) were cultured according to the providers’ guidelines.

Techniques: Derivative Assay, In Vitro, Staining, Angiogenesis Assay, Cell Culture, MANN-WHITNEY

Journal: bioRxiv

Article Title: A novel approach for reliable differentiation of lymphatic endothelial progenitor cells in vitro

doi: 10.1101/2025.09.23.677051

Figure Lengend Snippet: Stromal vascular fraction (SVF)-derived lymphatic endothelial progenitor cells (LEPCs) express canonical lymphatic markers in a 3D environment. Representative immunofluorescence staining images of LEPCs expressing lymphatic markers (PDPN, LYVE1, VEGFR3 and F-actin) embedded in Matrigel for 10 days. To the right, magnification of the white square in the left. Scale bar: 100 µm.

Article Snippet: Human dermal lymphatic endothelial cells (HDLEC, C-14021, PromoCell) were cultured according to the providers’ guidelines.

Techniques: Derivative Assay, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: A novel approach for reliable differentiation of lymphatic endothelial progenitor cells in vitro

doi: 10.1101/2025.09.23.677051

Figure Lengend Snippet: RNA sequencing analysis of Stromal vascular fraction (SVF)-derived lymphatic endothelial progenitor cells (LEPCs) confirm lymphatic genotype in comparison to mesenchymal progenitor cells. (A) Variance-stabilized transformed (VST) counts were used for Principal Component Analysis (PCA) plot. Axes show the top two principal components (PC1: 69.2%, PC2: 9.3%). Samples are colored by condition. (B) Sample clustering of the top 500 most variable genes. Distances between genes were estimated based on spearman correlation, which were then used to produce a clustering via the ward.D2 method. (C) Volcano plot of the differentially expressed genes (DEGs) between LEPCs and MSCs. Gray: non-significant; red: significant DEGs (|log2FC|≥1, adjusted p -value<0.05). (D) Boxplot of the differential expression of canonic lymphatic markers. Significance: ***adjusted p -value<0.001 (DESeq2 Wald test). (E) Heatmap of top 50 lymphatic endothelial cell (LEC) markers. Z-score-scaled VST counts for the 50 most significant LEC markers (rows) across samples (columns). Bold genes: |log2 fold change|≥1, adjusted p -value≤0.05. Clustering uses Euclidean distance and Ward.D2 linkage.

Article Snippet: Human dermal lymphatic endothelial cells (HDLEC, C-14021, PromoCell) were cultured according to the providers’ guidelines.

Techniques: RNA Sequencing, Derivative Assay, Comparison, Transformation Assay, Quantitative Proteomics