Journal: American Journal of Cancer Research

Article Title: Upregulation of Cyclin B2 ( CCNB2 ) in breast cancer contributes to the development of lymphovascular invasion

doi:

Figure Lengend Snippet: Representative photomicrographs of tumour cell adhesion across vascular and lymphatic endothelial cells (HUVECs, DLECs) (A) MCF-7, (B) ZR-75-1 cells and (C) SK-BR-3. (A-C) Silencing CCNB2 decreased the number of all BC cell lines adhered with HUVECs and DLECs. Representative photomicrographs of tumour cell (MCF-7, ZR-75-1 and SK-BR-3) transmigration across (D) HUVECs and (E) DLECs. (D, E) The number of tumour cells transmigrated across HUVECs and DLECs was higher in the control group compared to the KD group. Results shown are mean ± standard error of the mean (SEM) of three independent experiments. The P-values *<0.05, **<0.01, ***<0.001 and ****<0.0001.

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) and dermal lymphatic endothelial cells (DLECs) were purchased from Promocell (Heidelberg, Germany).

Techniques: Transmigration Assay

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

Journal: Pharmaceuticals

Article Title: Use of Early Donated COVID-19 Convalescent Plasma Is Optimal to Preserve the Integrity of Lymphatic Endothelial Cells

doi: 10.3390/ph15030365

Figure Lengend Snippet: Elevated antibody concentrations and prolonged symptoms are detrimental for the lymphatic endothelium integrity. ( A ) Correlation between the duration of symptoms (square-root transformation) and the concentration of plasmatic SARS-CoV-2- receptor-binding-domain antibodies measured by ELISA. ( B ) Correlation between the duration of symptoms and MitoSOX TM Red-negative cells measured by flow cytometry after incubating aHDLEC with convalescent plasma for 4 h and cytokines cocktail for 20 h. ( C ) Correlation between the concentration of plasmatic RBD antibodies measured by ELISA and the permeability of the endothelium measured by spectrophotometry (absorbance of OVA-488) after incubating human LEC with convalescent plasma for 4 h and cytokines cocktail for 20 h. Each point represents a treatment. Significance ( p < 0.05) was determined by a Pearson correlation. A square-root transformation of the duration of symptoms to reach normal distribution was performed. RBD—receptor-binding domain; O.D—optical density; OVA—ovalbumin; aHDLEC—adult human dermal lymphatic endothelial cells.

Article Snippet: Primary adult human dermal lymphatic endothelial cells (aHDLEC, PromoCell, cat. C-12217) were used in this study and called adult human lymphatic endothelial cells (aHDLEC) throughout the manuscript.

Techniques: Transformation Assay, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Permeability, Spectrophotometry

Journal: Pharmaceuticals

Article Title: Use of Early Donated COVID-19 Convalescent Plasma Is Optimal to Preserve the Integrity of Lymphatic Endothelial Cells

doi: 10.3390/ph15030365

Figure Lengend Snippet: The severity of symptoms correlates with the duration of symptoms, age, receptor-binding-domain antibodies of donors and cell viability of lymphatic endothelial cells. ( A ) Severity was quantified using a questionnaire answered by donors and graded as follows: asymptomatic, mild, moderate and severe. Severe symptoms were correlated with a higher duration of symptoms (square-root transformation to reach a normal distribution). ( B ) Correlations between severity of the symptoms and the age of the donors were performed. ( C ) Severity was correlated with the concentration of antibodies measured by ELISA. ( D , E ) Treated aHDLEC were labeled with Annexin V and PI and analyzed by flow cytometry. The severity of the symptoms was correlated with cells in late apoptosis/necrosis ( D ) and in early apoptosis ( E ) represented by Annexin V- and PI-positive cells and Annexin V-positive and PI-negative cells, respectively. Cells are given as percentages relative to cells treated with control plasma. Significance was determined by a Spearman correlation. p < 0.05 was considered significant. RBD, receptor-binding domain; O.D, optical density; PI, propidium iodide; aHDLEC, adult human dermal lymphatic endothelial cells.

Article Snippet: Primary adult human dermal lymphatic endothelial cells (aHDLEC, PromoCell, cat. C-12217) were used in this study and called adult human lymphatic endothelial cells (aHDLEC) throughout the manuscript.

Techniques: Binding Assay, Transformation Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Labeling, Flow Cytometry

Journal: Pharmaceuticals

Article Title: Use of Early Donated COVID-19 Convalescent Plasma Is Optimal to Preserve the Integrity of Lymphatic Endothelial Cells

doi: 10.3390/ph15030365

Figure Lengend Snippet: Early donation of COVID-19 convalescent plasma is a good predictor of preserved endothelial integrity. ( A , B ) Treated aHDLEC (convalescent plasma for 4 h and cytokines for 20 h) were harvested and the mRNA expression, measured by RT-qPCR, of SELE ( A ) and ICAM1 ( B ) was correlated with the time of donation since the onset of symptoms. ( C ) Following treatment, cells were fixed in paraformaldehyde (PFA) 2% and incubated with anti-VE-Cadherin antibodies. The intensity of VE-Cadherin relative to cells treated with control plasma was correlated to the duration between the onset of symptoms and the donation. ( D ) Immunofluorescence images of treated aHDLEC incubated with CCP donated at 27 days (upper panels) and 101 days (lower panels) post onset of symptoms. The left panels show the expression of VE-Cadherin and DAPI, whereas the middle panel represents the WGA staining. The right panels show the representation of the merged staining. Scale bar = 50 μm. ( E ) VE-Cadherin mRNA expression was correlated to the duration between the onset of symptoms and donation (∆). ( F ) Endothelial permeability was analyzed by the relative concentration of ovalbumin-488 measured following migration through the endothelium compared to control. The concentration of OVA-488 was correlated with the duration between the onset of symptoms and the donation. Significance was determined by Pearson correlation and p < 0.05 was considered significant. ∆, time of donation since onset of symptoms; SELE , gene coding for E-selectin; ICAM1 , gene coding for intercellular adhesion molecule 1; WGA, wheat germ agglutinin; CDH5 , gene coding for VE-Cadherin; OVA, ovalbumin; aHDLEC, adult human dermal lymphatic endothelial cells; CCP, COVID-19 convalescent plasma.

Article Snippet: Primary adult human dermal lymphatic endothelial cells (aHDLEC, PromoCell, cat. C-12217) were used in this study and called adult human lymphatic endothelial cells (aHDLEC) throughout the manuscript.

Techniques: Expressing, Quantitative RT-PCR, Incubation, Immunofluorescence, Staining, Permeability, Concentration Assay, Migration

Journal: Pharmaceuticals

Article Title: Use of Early Donated COVID-19 Convalescent Plasma Is Optimal to Preserve the Integrity of Lymphatic Endothelial Cells

doi: 10.3390/ph15030365

Figure Lengend Snippet: Secretion of lymphatic endothelial cells derived extracellular vesicles is a marker of impaired integrity. ( A ) Treated aHDLEC (convalescent plasma for 4 h and cytokines for 20 h) labeled by the MitoSOX TM Red probe and correlated with the quantity of EVs shed by the lymphatic endothelial cells. ( B , C ) Cells in sub-G 0 /G 1 ( B ) and G 2 /M ( C ) phases of the cell cycle, determined by cell cycle analysis using PI, were correlated to the quantity of EVs secreted by the lymphatic endothelium. ( D , E ). Treated aHDLEC were harvested and expression of FLT4 was assessed by RT-qPCR ( D ) and immunoblot ( E ) followed by a normalization onto the expression of ACTB or α/ß tubulin, respectively. The measured expression of VEGFR-3 was then correlated with the quantity of EVs secreted by the endothelium. ( F ) Immunoblot analysis for VEGFR-3 and the loading control α/ß tubulin. The left panel represents aHDLEC secreting low quantities of CD45 − podoplanin + EVs (less than 120,000 EVs) following incubation with CCP, and the right represents aHDLEC secreting high quantities of CD45 − podoplanin + EVs (more than 380,000 EVs). Significance was determined by a Pearson correlation and p < 0.05 was considered significant. EVs, extracellular vesicles; PI, propidium iodide; CCP, COVID-19 convalescent plasma; VEGFR-3, vascular endothelial growth factor receptor 3; FLT4, gene coding for VEGFR-3; aHDLEC, adult human dermal lymphatic endothelial cells; AU, arbitrary unit.

Article Snippet: Primary adult human dermal lymphatic endothelial cells (aHDLEC, PromoCell, cat. C-12217) were used in this study and called adult human lymphatic endothelial cells (aHDLEC) throughout the manuscript.

Techniques: Derivative Assay, Marker, Labeling, Cell Cycle Assay, Expressing, Quantitative RT-PCR, Western Blot, Incubation