ic 21 mouse macrophages  (ATCC)

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