cell growth medium 2  (PromoCell)

Journal: Journal of Extracellular Biology

Article Title: Proteomic Epithelial‐To‐Mesenchymal Transition Signature in Fetoplacental Small Extracellular Vesicles of Early‐Onset Preeclampsia

doi: 10.1002/jex2.70122

Figure Lengend Snippet: Characterisation of sEVs secreted by fetoplacental endothelial cells (fpECs) from term (T, n = 6), preterm (PT, n = 6) and Early‐Onset preeclamptic (EO‐PE, n = 4) pregnancies. (A) Representative transmission electron microscopy (TEM) images of sEVs (scale bar: 200 nm). (B) NTA: Size and concentration of particles, with size (x‐axis) plotted against particle concentration normalised to cell count at the time of isolation (y‐axis). Data are presented as mean ± standard error (SE). (C) Violin plots display the total concentration of sEVs (normalised to cell count at isolation) as measured by NTA. Statistical analysis was performed using the Kruskal–Wallis test, followed by pairwise Wilcoxon rank‐sum tests with Bonferroni correction for multiple comparisons. Adjusted p values are indicated (* = p < 0.05; ns = not significant). (D) Western blot analysis of sEVs using established sEV markers (Alix, TSG101, Syntenin‐1), endothelial marker (CD31) and non‐vesicular marker (ApoB).

Article Snippet: The cell pellet was resuspended in Endothelial Cell Growth Medium MV (ECGM; PromoCell, C‐22220, Heidelberg, Germany), supplemented with endothelial cell growth factor, hydrocortisone, epidermal growth factor (PromoCell, C‐39220), 0.05 mg/mL gentamicin (Gibco) and 10% defined FCS and plated in 12‐well plates coated with 1% gelatine (Sigma–Aldrich, St. Louis, MO).

Techniques: Transmission Assay, Electron Microscopy, Concentration Assay, Cell Characterization, Isolation, Western Blot, Marker

Journal: Journal of Extracellular Biology

Article Title: Proteomic Epithelial‐To‐Mesenchymal Transition Signature in Fetoplacental Small Extracellular Vesicles of Early‐Onset Preeclampsia

doi: 10.1002/jex2.70122

Figure Lengend Snippet: Descriptive statistics of proteomic analysis of fetoplacental‐endothelial derived sEVs from T ( n = 4), PT ( n = 5) and EO‐PE ( n = 4) pregnancies. (A) Venn diagram of identified proteins in the respective groups. Proteins were filtered to include those with >75% valid values in each group, resulting in 1098 proteins identified in T‐sEVs, 1179 proteins in PT‐sEVs and 1045 proteins in EO‐PE‐sEVs. (B) Principal component analysis (PCA) of protein abundances in the proteome. PCA revealed distinct clustering between the proteomes of EO‐PE‐ and PT‐sEVs, while T‐sEVs overlapped both, indicating shared variability. (C) Volcano plots of quantified proteins. Significant proteins are highlighted in dark grey (T), blue (PT) and red (EO‐PE) circles (two‐sided t ‐test with p values corrected for FDR, statistically significant if absolute log 2 FC ≥ 1.2 and p value ≤ 0.05), while non‐significant proteins are depicted in smaller grey circles.

Article Snippet: The cell pellet was resuspended in Endothelial Cell Growth Medium MV (ECGM; PromoCell, C‐22220, Heidelberg, Germany), supplemented with endothelial cell growth factor, hydrocortisone, epidermal growth factor (PromoCell, C‐39220), 0.05 mg/mL gentamicin (Gibco) and 10% defined FCS and plated in 12‐well plates coated with 1% gelatine (Sigma–Aldrich, St. Louis, MO).

Techniques: Derivative Assay