ccd camera  (Bio-Rad)

Journal: Journal of Extracellular Vesicles

Article Title: Lipidome Analysis of Cancer Cells and Their Extracellular Vesicles Reveals Cancer‐Type‐Specific Lipid Signatures and Enables the Design of EV‐Mimetic Liposomes

doi: 10.1002/jev2.70265

Figure Lengend Snippet: Characterization of EVs derived from different cell lines. Figure presents the strategy for isolating and characterizing EVs from six human cell models, illustrating their morphological diversity, homogeneous size (130–160 nm), negative surface charge, and expression of the syntenin marker. These results validate the quality and comparability of the EVs used for lipidomic analysis. (A) Schematic representation of the cell lines used for EV isolation. Created in https://BioRender.com . (B) TEM images of EVs isolated from MP41 (primary tumor), OMM2.5 (metastatic tumor) and BJ (non‐cancer) cells, showing the spherical morphology and nanometric size of EVs. Scale bars: 500 nm. (C) Normalized distribution (min–max normalization) of EV size, measured by NTA. The Y ‐axis values represent relative proportions between 0 and 1, calculated based on the minima and maxima of each sample. This representation allows for comparison of the shapes of the distributions regardless of differences in initial particle concentration. (D) Mean size (in nm) of EVs measured by NTA. (E) Zeta potential of EVs measured in millivolts (mV), indicating the surface charge of the particles. The zeta potential was negative for all EV samples, as expected. (F) Reconstituted western blots showing the presence of syntenin (detected at ∼35 kDa) in EVs isolated from four cancer cell lines (HT29, MP41, MEL270 and OMM2.5) and two non‐cancer cell lines (BJ, CCD‐18Co). The antibody was used at a 1:1,000 dilution. All experiments were performed in triplicates. Note: syntenin was selected as an EV marker due to its abundance and conservation in small EVs, in line with recent proteomic studies and MISEV2023 guidelines. Furthermore, our previous work (Lopez et al. ; Tsering et al. ) confirmed the presence of other canonical EV markers, including TSG101 and CD81, in a subset of these cell lines.

Article Snippet: CCD‐18Co , EMEM (Corning) supplemented with 10% FBS and 0.1% 10 U/mL penicillin and 10 μg/streptomycin , American Type Culture Collection (ATCC).

Techniques: Derivative Assay, Expressing, Marker, Isolation, Comparison, Concentration Assay, Zeta Potential Analyzer, Western Blot

Journal: Journal of Extracellular Vesicles

Article Title: Lipidome Analysis of Cancer Cells and Their Extracellular Vesicles Reveals Cancer‐Type‐Specific Lipid Signatures and Enables the Design of EV‐Mimetic Liposomes

doi: 10.1002/jev2.70265

Figure Lengend Snippet: Individual lipidomic profiling of cells and their EVs. Figure highlights the diversity of lipid families and saturation levels in each cell line and their EVs. The profiles reveal distinct lipid signatures depending on the cell type, with a predominance of phospholipids and marked differences between cancerous and non‐cancerous cells. (A) Schematic representation that presents the type of analysis (a descriptive characterization of the lipid profile of each sample) and key information. Created in https://BioRender.com . (B) Pie charts showing the relative proportions of lipid classes (lysophospholipids, phospholipids, sphingolipids and sterols) for each cell line and their EVs: MP41, MEL270, OMM2.5, HT29, CCD‐18Co and BJ. (C) Line graphs depict the relative abundance of individual lipid species within each family for cells and EVs from each sample type. The X ‐axis represents lipid families, while the Y ‐axis indicates the percentage of lipid species. (D) Pie charts showing the overall proportions of saturated, monounsaturated and polyunsaturated lipids in cells and EVs for each sample.

Article Snippet: CCD‐18Co , EMEM (Corning) supplemented with 10% FBS and 0.1% 10 U/mL penicillin and 10 μg/streptomycin , American Type Culture Collection (ATCC).

Techniques:

Journal: Journal of Extracellular Vesicles

Article Title: Lipidome Analysis of Cancer Cells and Their Extracellular Vesicles Reveals Cancer‐Type‐Specific Lipid Signatures and Enables the Design of EV‐Mimetic Liposomes

doi: 10.1002/jev2.70265

Figure Lengend Snippet: Cell‐EV lipid profile comparison. Figure compares the lipid profiles of cells and their EVs, showing that nearly half of the lipid species are shared, but that each cell type exhibits specific enrichments or depletions in its EVs. These differences highlight lipid sorting mechanisms specific to each cellular context. (A) Schematic representation that presents the type of analysis (cell‐EV lipid profile comparison) and key information. Created in https://BioRender.com . (B) Venn diagrams illustrating the overlap of identified lipid species between cells and EVs for each sample type: MP41, MEL270, OMM2.5, HT29, CCD‐18Co and BJ. The numbers represent the total lipid species unique to cells, unique to EVs, and shared between both. (C) Bar graphs summarize the proportions of different lipid families in EVs for each cell line. The Y ‐axis represents lipid families, while the X ‐axis indicates the percentage of lipid species in EVs, compared to their cells. (D,E) Volcano plots showing the differential abundance of lipid species shared between cells and their corresponding EVs. Panel (D) Highlights the underexpressed (green) and overexpressed (red) lipids in cells compared to their EVs, while panel (E) Presents the underexpressed (green) and overexpressed (red) lipids in EVs compared to their corresponding cells. The X ‐axis represents the log 2 fold change (log 2 FC), while the Y ‐axis indicates statistical significance (−log 10 p ‐value). Dashed lines indicate significance thresholds ( p < 0.05). (F,G) Pie charts display the relative proportions of lipid classes (lysophospholipids, phospholipids, sphingolipids and sterols) among the lipids identified as significantly up‐regulated in cells (F) or in EVs (G) based on panels (D) and (E) . These charts provide a focused view of the lipid family composition within the subsets of over‐expressed lipids.

Article Snippet: CCD‐18Co , EMEM (Corning) supplemented with 10% FBS and 0.1% 10 U/mL penicillin and 10 μg/streptomycin , American Type Culture Collection (ATCC).

Techniques: Comparison

Journal: Journal of Extracellular Vesicles

Article Title: Lipidome Analysis of Cancer Cells and Their Extracellular Vesicles Reveals Cancer‐Type‐Specific Lipid Signatures and Enables the Design of EV‐Mimetic Liposomes

doi: 10.1002/jev2.70265

Figure Lengend Snippet: Global comparison of lipid profiles across all samples. Figure shows, through clustering and heatmap analyses, a clear separation between cells and EVs, as well as between cancer and non‐cancer models. Each line and its EVs exhibit unique lipid signatures, illustrating the specific metabolic adaptation to each biological context. Brown box (cells and EVs) : (A) Schematic representation that presents the analysis of all samples. Created in https://BioRender.com . (B) PCA illustrating the distribution of all the samples based on their lipidomic profiles. Each point represents a sample, with clustering reflecting the overall lipid composition of cells and EVs. Orange box (cells) : (C) Schematic representation that presents the cell analysis and key information. Created in https://BioRender.com . ( D) PCA plot showing the separation of cancer cells (MP41, MEL270, OMM2.5 and HT29) and non‐cancer cells (CCD‐18Co, BJ) based on lipid profiles. Each point represents a sample, and clustering reflects differences in lipid composition. (E) Venn diagram showing the overlap of lipid species identified in the six cell lines: MP41, MEL270, OMM2.5, HT29 and CCD‐18Co, and BJ. Numbers indicate lipid species unique to each cell line and those shared across multiple cell types. (F) Heatmaps depicting the relative abundance of lipid species shared among the six cell lines. Green indicates lower abundance, while red indicates higher abundance. (G) Pie charts illustrating the relative proportions of lipid classes (lysophospholipids, phospholipids, sphingolipids and sterols) and saturation levels (saturated, monounsaturated and polyunsaturated lipids) among the unique lipid species identified in each cell line. (H) Lists of unique lipids in cells. Blue box (EVs) : (I) Schematic representation that presents the EV analysis and key information. Created in https://BioRender.com . (J) PCA plot showing the separation of EVs derived from cancer cells and non‐cancer cells based on lipid profiles. Each point represents a sample, with clustering indicating compositional differences. (K) Venn diagram showing the overlap of lipid species identified in EVs derived from the six cell lines. Numbers indicate lipid species unique to each EV sample and those shared across multiple EV types. (L) Heatmaps depicting the relative abundance of lipid species shared among EVs from the six cell lines. Green indicates lower abundance, while red indicates higher abundance. (M) Pie charts illustrate the relative proportions of lipid classes and saturation levels among the unique lipid species identified in EVs derived from each cell line. (N) Lists of unique lipids in EVs.

Article Snippet: CCD‐18Co , EMEM (Corning) supplemented with 10% FBS and 0.1% 10 U/mL penicillin and 10 μg/streptomycin , American Type Culture Collection (ATCC).

Techniques: Comparison, Cell Analysis, Derivative Assay