Journal: Theranostics

Article Title: Toward the clinical translation of safe intravenous long circulating ILNEs contrast agent for CT imaging.

doi: 10.7150/thno.110014

Figure Lengend Snippet: Figure 3. Characterization and stability of ILNEs in various conditions. (A-B) Particle size stability in HP and (C-D) FBS: 10% and 20% (v/v) ILNEs were incubated with (A-B) HP or FBS (C-D) for 48 h. Particle size (A-C) and PDI (B-D) were measured at different time points using DLS. (E) UV-vis absorbance measurements of ILNE1–4, showing the peaks of CrEL and TIPhO at λmax of ~230 nm and ~244 nm, respectively. (F) Viscosity measurements of ILNE1–4, compared with iodixanol stock solution (300 mgI/mL) at 60% (wt/v). (G) Phantom scan of ILNEs: In vitro evaluation of x-ray attenuation properties (in HU) of ILNE1–4 using a calibration curve made by various dilutions of the iodixanol injection as a reference (expressed in mgI/mL). Air and water were used for normalization. Top view of Eppendorf tubes and source data are provided as a Source Data file. (H) CCK-8 cytotoxicity assay of immune cells: Macrophages RAW264.7 and IC21 cells were incubated with ILNE3 for 48 h. Data are presented as the average ± SD (n = 6). (I) Cell viability studies of ILNE components: Two concentrations of CrEL and TIPhO were incubated with RAW 264.7 cells for 24 h.

Article Snippet: Human Dermal Fibroblasts, neonatal (HDFn), RAW 264.7 macrophages and IC21 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Incubation, Viscosity, In Vitro, Injection, CCK-8 Assay, Cytotoxicity Assay

Journal: Scientific reports

Article Title: Cationized extracellular vesicles for gene delivery.

doi: 10.1038/s41598-024-75985-y

Figure Lengend Snippet: Fig. 6. Transfection of IC21 macrophages with MVL5-EVs-based formulations of pDNA. (A) MVL5-EVs (20 µg MVL5/1011particles) were prepared via MVL5 incorporation into EVs by sonication at different temperatures (PEG purification after MVL5 incorporation), and then loaded with luciferase pDNA (6 µg /1011particles). IC21 macrophages (1 × 105 cells/well) were supplemented with MVL5-EVs formulations of pDNA (1 × 1010 particles/1 µg pDNA/well) and incubated for 4 h at 37 °C. (B) MVL5-EVs were prepared via incorporation of different amounts of MVL5 (0.5–50 µg MVL5/1011particles) into EVs by sonication in the presence or absence of EtOH (50%) and added to IC21 macrophages (2 × 104 cells/well). Free pDNA (1 µg/ well), or mixture of EVs and pDNA (1010particles/1 µg pDNA/well), or mixture of common transfection agent, GenePorter (GP3K, according to manufacturer’s protocol), with pDNA (1 µg/well) at the same concentration were used as controls. Then, the cells were washed with PBS and cultured in full media for 24 h, and the luminescence of lysed cells in the presence of luciferin-ATP was measured. White bars – samples without EVs. (A) Bell shaped curve was observed for transfection efficacy with maximum at 42 °C used for cationized EVs preparation. (B) Transfection efficacy increased with the increases of MVL5 incorporated into EVs nanocarriers. Little, if any, luminescence was detected in cells exposed to naked pDNA, or mixture pDNA with non-cationized EVs. As expected, significant transfection was observed in the cells supplemented mixture of free pDNA and the cationic lipid, or free pDNA and GP3K.

Article Snippet: Mouse macrophage cell lines (Raw 264.7, and IC21), and TNBC cells MDA-MB 231, were purchased from ATCC (Manassas, VA, USA).

Techniques: Transfection, Sonication, Purification, Luciferase, Incubation, Concentration Assay, Cell Culture

Journal: Scientific reports

Article Title: Cationized extracellular vesicles for gene delivery.

doi: 10.1038/s41598-024-75985-y

Figure Lengend Snippet: Fig. 8. Transfection IC21 macrophages with MVL5-EVs-luc-mRNA. MVL5-EVs (20 µg MVL5/1011particles) were prepared via MVL5 incorporation into EVs by sonication at 42 °C with PEG purification after MVL5 incorporation, and then loaded with luciferase mRNA (1.3 µg /1011 particles). IC21 macrophages (1 × 105 cells/ well) were supplemented with MVL5-EVs-Luc-mRNA (3 µg/1011 particles/well) loaded with different amount of mRNA (0.4–3 µg) and incubated for 4 h at 37 °C. Naked mRNA (maximum amount, 3 µg/well) with or without GP3K was used as a control. Then, the cells were washed with PBS and cultured in full media for 24 h, and the luminescence of lysed cells in the presence of luciferin-ATP was measured. Cationized EVs provided efficient transfection in vitro. MVL5-EVs were obtained by lipid incorporation using sonication in water bath followed by PEG precipitation.

Article Snippet: Mouse macrophage cell lines (Raw 264.7, and IC21), and TNBC cells MDA-MB 231, were purchased from ATCC (Manassas, VA, USA).

Techniques: Transfection, Sonication, Purification, Luciferase, Incubation, Control, Cell Culture, In Vitro