Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Paper-Based Devices for Capturing Exosomes and Exosomal Nucleic Acids From Biological Samples

doi: 10.3389/fbioe.2022.836082

Figure Lengend Snippet: The scheme describes the experimental design in this study. HCT116 cells were cultured in various microenvironments (pH 7.4 and 6.5), characterized by microscope, ultrafiltrated to collect concentrated exosomes, which were then analyzed by a qNano instrument to determine their concentration and size distribution, and subjected to paper-based immunoaffinity devices for exosome capture. The morphology of captured exosomes was subsequently characterized by SEM and their amount was quantified by ELISA. Eventually, exosomal nucleic acids were absorbed by paper-based silica devices for miR-21 quantification by RT-qPCR.

Article Snippet: On the one hand, concentrations of 3.6×10 8 exosomes/mL and size distributions of approximately 110–160 nm (mean diameter of 133.1 nm, ) were observed, which was equivalent in amount and identical in size to those of the standard sample (3.4×10 9 exosomes/mL and 100–200 nm (mean diameter of 129 nm, ) identified from a 1.9×10 9 exosome/mL solution of commercially available lyophilized exosomes from the HCT116 cell line (human colon carcinoma) obtained from Hansa BioMed Life Science, Estonia), indicating that HCT116 cell-derived exosomes pretreated by our proposed method met the commercial requirements.

Techniques: Cell Culture, Microscopy, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Paper-Based Devices for Capturing Exosomes and Exosomal Nucleic Acids From Biological Samples

doi: 10.3389/fbioe.2022.836082

Figure Lengend Snippet: Particle size distribution and concentration of (A) exosomes from HCT116 cell culture medium and (B) commercial lyophilized exosomes analyzed by qNano.

Article Snippet: On the one hand, concentrations of 3.6×10 8 exosomes/mL and size distributions of approximately 110–160 nm (mean diameter of 133.1 nm, ) were observed, which was equivalent in amount and identical in size to those of the standard sample (3.4×10 9 exosomes/mL and 100–200 nm (mean diameter of 129 nm, ) identified from a 1.9×10 9 exosome/mL solution of commercially available lyophilized exosomes from the HCT116 cell line (human colon carcinoma) obtained from Hansa BioMed Life Science, Estonia), indicating that HCT116 cell-derived exosomes pretreated by our proposed method met the commercial requirements.

Techniques: Concentration Assay, Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Paper-Based Devices for Capturing Exosomes and Exosomal Nucleic Acids From Biological Samples

doi: 10.3389/fbioe.2022.836082

Figure Lengend Snippet: SEM pictures characterize surface morphology of (A) untreated sample, (B) commercial lyophilized exosomes, and (C,D) HCT116-derived exosomes captured by paper-based immunoaffinity devices. Red arrows indicate captured exosomes.

Article Snippet: On the one hand, concentrations of 3.6×10 8 exosomes/mL and size distributions of approximately 110–160 nm (mean diameter of 133.1 nm, ) were observed, which was equivalent in amount and identical in size to those of the standard sample (3.4×10 9 exosomes/mL and 100–200 nm (mean diameter of 129 nm, ) identified from a 1.9×10 9 exosome/mL solution of commercially available lyophilized exosomes from the HCT116 cell line (human colon carcinoma) obtained from Hansa BioMed Life Science, Estonia), indicating that HCT116 cell-derived exosomes pretreated by our proposed method met the commercial requirements.

Techniques: Derivative Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Paper-Based Devices for Capturing Exosomes and Exosomal Nucleic Acids From Biological Samples

doi: 10.3389/fbioe.2022.836082

Figure Lengend Snippet: Quantification of exosomes performed by color intensity assessment including commercial lyophilized samples (blue columns) and exosomes captured from the medium of HCT116 cells cultured in various microenvironments of pH 7.4 (red column) and 6.5 (green column). Inset is the calibration curve of exosomes from commercial samples (blue columns). An asterisk (*) denotes a p value <0.05 (obtained from t -test). All of the results were obtained by P-ELISA.

Article Snippet: On the one hand, concentrations of 3.6×10 8 exosomes/mL and size distributions of approximately 110–160 nm (mean diameter of 133.1 nm, ) were observed, which was equivalent in amount and identical in size to those of the standard sample (3.4×10 9 exosomes/mL and 100–200 nm (mean diameter of 129 nm, ) identified from a 1.9×10 9 exosome/mL solution of commercially available lyophilized exosomes from the HCT116 cell line (human colon carcinoma) obtained from Hansa BioMed Life Science, Estonia), indicating that HCT116 cell-derived exosomes pretreated by our proposed method met the commercial requirements.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Paper-Based Devices for Capturing Exosomes and Exosomal Nucleic Acids From Biological Samples

doi: 10.3389/fbioe.2022.836082

Figure Lengend Snippet: Quantification of miR-21. (A) Calibration curve for standard miR-21 samples by paper-based silica devices and quantified by RT-qPCR. (B) miR-21 contents (estimated by Ct values) in exosomes from standard samples and from 10 to 20 ml pH HCT116 samples cultured in pH 7.4 and pH 6.5 microenvironments. Three asterisks (***) denote a p value <0.001 (obtained from t -test). (C) miR-21 concentrations (copies/µL) of 10 and 20 ml HCT116 samples cultured in pH 7.4 and pH 6.5 microenvironments, calculated from calibration line in (A) and corresponding Ct values in (B) . (D) U6 SiRNA contents (estimated by Ct values) derived from HCT116 cells cultured in pH 7.4 (red column) and pH 6.5 (green column) conditions.

Article Snippet: On the one hand, concentrations of 3.6×10 8 exosomes/mL and size distributions of approximately 110–160 nm (mean diameter of 133.1 nm, ) were observed, which was equivalent in amount and identical in size to those of the standard sample (3.4×10 9 exosomes/mL and 100–200 nm (mean diameter of 129 nm, ) identified from a 1.9×10 9 exosome/mL solution of commercially available lyophilized exosomes from the HCT116 cell line (human colon carcinoma) obtained from Hansa BioMed Life Science, Estonia), indicating that HCT116 cell-derived exosomes pretreated by our proposed method met the commercial requirements.

Techniques: Quantitative RT-PCR, Cell Culture, Derivative Assay