Journal: Biomaterials science

Article Title: Matrix stiffness and stress relaxation regulate matrix-bound nanovesicle release from alginate hydrogels

doi: 10.1039/d6bm00832a

Figure Lengend Snippet: (A) Schematic depiction of MBV isolation procedure from in vitro dermal fibroblast cultures. (B) Nanoparticle tracking analysis of diluted MBVs (1:10,000) measures particle size distribution and concentration. (C) Transmission electron micrograph of MBVs reveals vesicle-like morphology. (D) Western blot analysis of Alix (~100kDa), Integrin β1 (~100–130 kDa), CD63 (~35–55 kDa), and GRP94 (~100 kDa) expression in MBVs and dermal fibroblast cell lysate. (E) Fluorescence and scanning electron microscopy images of fast-relaxing and slow-relaxing alginate hydrogels with and without MBVs. Top: 5kPa hydrogels encapsulating CFSE-labeled MBVs, * denotes region outside of hydrogel. Bottom: 20kPa hydrogels encapsulating unlabeled MBVs.

Article Snippet: Primary antibodies were diluted in 5% w/v bovine serum albumin in 1X PBS-T (0.2%), including anti-Integrin β1 (1:1,000, Cell Signaling Technology, 34971), anti-Alix (1:10,000, Proteintech, 12422–1-AP), anti-CD63 (1:1,000, Proteintech, 25682–1-AP), and GRP94 (1:5,000, Proteintech, 14700–1-AP).

Techniques: Isolation, Encapsulation, In Vitro, Analysis, Concentration Assay, Transmission Assay, Morphology, Western Blot, Expressing, Fluorescence, Scanning Electron Microscopy, Labeling, Cell Culture

Journal: Medicine

Article Title: Human amniotic epithelial cell-derived exosomes promote conjunctival goblet cells proliferation and mucin secretion

doi: 10.1097/MD.0000000000049647

Figure Lengend Snippet: Identification and characterization of hAECs-Exo. (A) Morphology of exosomes in the hAECs-Exo group was observed under transmission electron microscopy (Scalar bar = 200 nm). (B) Diameter of exosomes in the hAECs-Exo group was analysis under NTA. (C) Western blot was used to observe the expression of CD63, CD81, and Grp94 proteins in hAECs-Exo. hAECs-Exo = human amniotic epithelial cell-derived exosomes, NTA = nanoparticle tracking analysis.

Article Snippet: The polyvinylidene fluoride membrane was treated with protein-free rapid blocking solution (PS108P, EpiZyme) for 15 minutes, and then incubated with primary antibodies against exosome markers CD63 (1:1000, ABclonal, A19023) and CD81 (1:1000, ABclonal, A4863), as well as the negative marker Grp94 (1:1000, ABclonal, A0989) overnight.

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Expressing, Derivative Assay