Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Adipose Mesenchymal Cells-Derived EVs Alleviate DOCA-Salt-Induced Hypertension by Promoting Cardio-Renal Protection

doi: 10.1016/j.omtm.2019.11.002

Figure Lengend Snippet: ASC-EV Treatment Promoted Important Attenuation of Kidney Fibrosis in the DOCA-Salt Model (A) Representative photomicrographs of kidney slices stained with Picrosirius red on 9-week-treated rats from the 4 experimental groups (scale bars, 50 μm). (B) α-SMA expression in the kidney of 9-week-treated rats under the different experimental conditions was determined by immunohistochemistry (n = 4 for each treatment) (scale bars, 50 μm). (C) Quantification of Picrosirius red stained area in kidney sections expressed as a percentage of the total area of the kidney, in 9-week-treated rats, measured for all the animals tested in the 4 experimental groups (SHAM, n = 4; UNX, n = 9; DOCA, n = 13; DOCA-EV, n = 10). (D−F) Western blot analysis of α-SMA and Desmin in the cortex of 9-week-treated UNX (n = 6), DOCA, and DOCA-EV (n = 8) rats. GAPDH was used as loading control. Values represent the fold increase with respect to UNX (considered as the reference = 1). (G−I) Analysis of COL1A1 , COL4A1 , and FN mRNA levels in the renal tissue in control SHAM (n = 3) and in all the uninephrectomized animal groups (UNX, n = 6; DOCA and DOCA-EV, n = 7). qRT-PCR data were expressed as RQ and UNX was used as the reference sample. Data represent mean ± SE. *indicates statistical difference to SHAM group, # indicates statistical difference to UNX group, and § indicates statistical difference to DOCA group; p < 0.05.

Article Snippet: The following primary antibodies were used: GAPDH, sc-32233 (1:1,000, Santa Cruz); α-SMA, MAB1420 (1:1,000, R&D Systems, Minneapolis, MN, USA); Desmin, ab32362 (1:1,000, Abcam); B cell lymphoma 2 (Bcl2), 2876 (1:1,000, Cell Signaling Technology, Danvers, MA, USA); and Bcl-2-associated X (Bax), 2772S (1:500, Cell Signaling Technology).

Techniques: Staining, Expressing, Immunohistochemistry, Western Blot, Control, Quantitative RT-PCR

Journal: Biomaterials

Article Title: Fibrin polymer on the surface of biomaterial implants drives the foreign body reaction.

doi: 10.1016/j.biomaterials.2021.121087

Figure Lengend Snippet: Fig. 9. Macrophages within the fibrous capsule express α-SMA and CD68. PCTFE sections were implanted in WT (A, B) and FibAEK (C, D) mice for 3, 7, and 14 days. Representative images of the capsule sam ples retrieved from WT (n = 3) and FibAEK

Article Snippet: The mouse anti-α-SMA mAb (catalog number MAB1420-SP) was from R&D systems.

Techniques:

Journal: Biomaterials

Article Title: Fibrin polymer on the surface of biomaterial implants drives the foreign body reaction.

doi: 10.1016/j.biomaterials.2021.121087

Figure Lengend Snippet: Fig. 10. Macrophages within the fibrous capsule express collagen. PCTFE sections were implanted in WT and FibAEK mice for 14 days and the cells accumulated within the capsules were isolated as described in the Materials and Methods. (A) The cells were allowed to adhere to the surface of a FluoroDish, fixed, and incubated with Alexa Fluor 568-conjugated phalloidin, anti-α-SMA, and anti-CD68 antibodies. Representative confocal images of myofibroblasts and macrophages isolated from the capsule retrieved from WT mice are shown. Arrowheads point to podosomes seen in macrophages incubated with phalloidin-Alexa Fluor 568. The scale bar is 10 μm. (B) Quantification of cells expressing α-SMA only (myofibroblasts) or both α-SMA and CD68 (macrophages). Approximately 2000–2500 cells isolated from the capsules retrieved from each WT and FibAEK mice were analyzed with ~400 cells present in each field of view. (C) The cells isolated from the capsule obtained from WT mice were incubated with anti-CD11b mAb M1/70 and anti-collagen I mAb. Myofibroblasts shown in the upper panel are indicated by white arrows. The scale bar is 20 μm. (D) Representative confocal images of collagen-expressing macrophages isolated from the WT and FibAEK mice. The cells were labeled for collagen 1 (green) and actin (red). The scale bar is 10 μm. (E) The frequency distribution of fluorescence intensities for collagen I in macrophages isolated from the capsules retrieved from WT and FibAEK mice was expressed as CTCF arbitrary units (A.U). A total of 300 cells from 5 random fields in the samples prepared from WT and FgAEK

Article Snippet: The mouse anti-α-SMA mAb (catalog number MAB1420-SP) was from R&D systems.

Techniques: Capsules, Isolation, Incubation, Expressing, Labeling, Fluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: FACS-purified myoblasts producing controlled VEGF levels induce safe and stable angiogenesis in chronic hind limb ischemia

doi: 10.1111/j.1582-4934.2011.01308.x

Figure Lengend Snippet: FACS-purified rVICD8 myoblasts induce normal, stable and mature angiogenesis in ischemic muscle. (A–E) The medial hamstring muscles were harvested 3 months after myoblast injection and frozen sections were immunostained with antibodies against endothelium (PECAM, in red), pericytes (NG2, in green) and smooth muscle cells (α-SMA, in blue). Images were taken in areas where myoblast engraftment had been confirmed in adjacent serial sections stained with X-Gal. (A) nonischemic, non-treated muscle. (B) rICD8 control cells. (C) reference clone. (D) primary transduced rVICD8 population. (E) FACS-purified rVICD8 myoblasts. The white arrow in (D) indicates a large angioma-like aberrant structure. Size bars 50 7μm. (F) Vessel length density (VLD) was measured around engrafted fibres and is expressed as the mean vessel length (in m) per fibre. Values represent the mean ± S.E.M. ( n = 3–5). CD8: control cells; VICD8: primary transduced rVICD8 myoblasts. *P < 0.05 compared to control cells.

Article Snippet: The following primary antibodies and dilutions used were: mouse monoclonal anti-rat Platelet-Endothelial Cell Adhesion Molecule-1 (PECAM-1, Clone TLD-3A12; AbD Serotec, Düsseldorf, Germany) at 1:100; mouse monoclonal anti-α-SMA (Clone 1A4; MP Biomedicals, Basel, Switzerland) at 1:400; rabbit polyclonal anti-Nerve/Glial antigen-2 (NG2; Chemicon Europe, Chandlers Ford, UK) at 1:200.

Techniques: Purification, Injection, Staining