ALT Vesiculab webinar focusing on engineered extracellular vesicles, broadcasting on 18th September 2024
ALT Isolation and characterization of EVs from Salvia sclarea HR conditioned medium. (A) Representative image of S. sclarea HR culture employed as EV biofactory. (B) Conditioned media were collected from two-month-old HRs and used for the purification of EVs by differential ultracentrifugation (dUC) or size exclusion chromatography (SEC). (C) Protein profile analysis of HR-derived EVs visualized through SDS-PAGE and silver staining. (D and E) Nanoparticle Tracking Analysis (NTA) measurements depict the size distribution of S. sclarea HR EVs purified by dUC (dilution= 1: 100) and SEC (dilution= 1:5), respectively. (F) Scanning electron image showing a group of S. sclarea HR EVs. (G) and (H) Transmission electron microscopy (TEM) close-up images displaying the round-shaped morphology of S. sclarea HR EVs. (I) Amplification of rolB, rolC and virD2 genes, located in the pRi15834 agrobacterium-type Ri plasmid was carried out through PCR in HR EVs. pRi15834 plasmid extracted from A.
ALT Expanded USC express MSC-associated markers CD73, CD90, and CD105. After culturing, the expression of selected surface markers on clones collected at different time points was evaluated by flow cytometry. Stratification for gender is not included in the displayed results. The percentwise expression (A) and median fluorescence intensity (MFI) (B) for each surface marker, are provided as mean ± SEM for all included donors. (C) Representative histograms for the investigated surface marker as well as forward scatter (FSC) and side scatter (SSC)
ALT Extracellular vesicle miRNAs modulate glioblastoma TME. EV containing miRNAs released by glioblastoma cells target cellular proteins and influence the signaling pathways to modulate various cellular processes in donor and recipient cells such as resistance development, invasion, migration, tumour proliferation, GSC development, angiogenesis and immune suppression. The crosstalk and exchange of miRNAs between donor and recipient cells promote glioblastoma pathogenesis.
ALT Top image: After 6 h of cell exposure to the hUSC-EX, the immunofluorescence images showed hUSC-EX was uptaken and internalized by the injured ACL cells. ACL: anterior cruciate ligament; hUSC-EX: human umbilical cord stem cells derived exosome Bottom image: The expression of Collagen synthesis, TGFβ, and VEGF gene difference between the non-exosome treated 8 weeks injured ACL cells (control) and hUSC-EX treated cells after 24 h treatment. hUSC-EX: human umbilical cord stem cells derived exosome
ALT EV purified from in vivo mice model of HSV-1 infection. (A and B) Representative images of TEM (A) and ScEM (B) analyses showing the EV isolated from in vivo samples. Scale bars are indicated in the figure. (C) Size distribution of collected vesicle semi-quantitative data was obtained from TEM images measuring the major diameter of more than 150 EV. (D) Representative immunoblots showing tau phosphorylation and protein levels in lysates of different fractions of EV purification from mock- (ctr) and HSV-1-infected mice. Alix expression level was used as sample loading control. (E) Densitometric analyses of immunoreactive signals in Fraction 1 (F1) are shown in the graphs: values represent the normalized fold-changes in protein levels from HSV-infected mice with respect to ctr (n = 5). Data are expressed as mean ± SEM. Statistical significance was calculated by one-sample t-test.
ALT Efficient EV-mediated delivery of the immunomodulatory molecule mOx40L showed significant therapeutic impact in murine tumour model in vivo.
ALT Development of complementary analytical methods to characterize extracellular vesicles graphical abstract
ALT Top image: Tumor exosomal regulates cancer progression, like proliferation, invasion, metastasis, angiogenesis, and immunosuppression. Bottom image: The function of exosome noncoding RNAs in cancer. Tumor cells secrete exosomes, which contain diverse nucleic acids (ncRNA) to modify tumors cells and stromal cells in the tumor progression such as endothelial cells to effect angiogenesis, tumor cells develop resistance to drugs, growth metastasis, and immune disorder.
ALT Semi quantification of human serum derived AnnV+CD14+ large EVs. (a) Workflow SOP for isolation of large EVs (lEVs) from 1 mL of human serum collected from polytrauma patients (ISS > 15, at day 0, at day 1 and day 7 post trauma). (b) AnnV+CD14+ large EVs counts in small EVs in polytrauma (ISS > 15) with internal organ damage (polytrauma OD) versus polytrauma (ISS ≥ 15) without internal organ damage (polytrauma w/o OD) upon admission (day 0). (c) AnnV+CD14+ large EVs counts in polytrauma (ISS > 15) polytrauma OD versus polytrauma w/o OD (ISS > 15) 24 h post trauma (day 1). (d) Day 1, corresponding AUROC, sensitivity, specificity, and cut-off value, respectively. (e) AnnV+CD14+ large EVs counts in small EVs in polytrauma (ISS ≥ 15) polytrauma OD versus polytrauma w/o OD (ISS ≥ 15) 7 days post trauma (day 7). Values given as median with 95% CI.
ALT Top image: Roles of EVs in different female reproductive tissues. EVs differ in their molecular content and role in biological processes based on cell of origin and its environment. Figure was made using Biorender, adapted from Kurian and Modi (2019) and Qamar et al. (2020). Bottom image: Bioengineering platforms for improved EV production. The figure illustrates diverse bioengineering approaches employed in the in vitro production of extracellular vesicles (EVs), showcasing the continuum between throughput and physiological relevance. Each technology is strategically positioned on this spectrum to highlight its characteristics in terms of scalability and mimicry of natural physiological conditions. Figure was made using Biorender.