fibroblasts migration  (Exosome Diagnostics)

Journal: Journal of Translational Medicine

Article Title: Exosomes released from human induced pluripotent stem cells-derived MSCs facilitate cutaneous wound healing by promoting collagen synthesis and angiogenesis

doi: 10.1186/s12967-015-0417-0

Figure Lengend Snippet: Characterization of human induced pluripotent stem cell-derived mesenchymal stem cells (hiPSC-MSCs) and hiPSC-MSC-derived exosomes (hiPSC-MSC-Exos). (A) Light microscopy images demonstrating morphological changes occurring during hiPSCs differentiation into fibroblast-like cells. (a) Representative cell morphology of hiPSCs before differentiation. (b) Intermediate phase of differentiating the hiPSCs into MSCs. (c) Typical fibroblast-like morphology of cells. (B) Flow cytometric analysis of the surface markers in hiPSC-MSCs. (C) Assessment of the tri-lineage differentiation capacity of iPSC-MSC-like cells. (a) Alizarin Red staining for osteocytes after 3 weeks in culture with osteogenic medium. (b) Alcian Blue staining for chondrocytes after 4 weeks in culture with chondrogenic medium. (c) Oil Red O staining for adipocytes after 2 weeks in culture with adipogenic medium. The qRT-PCR results for OCN (d) , Sox9 (e) , and LPL (f) after 7 days in culture with osteo-, chondro-, and adipogenic mediun. (D) Transmission electron microscope images of hiPSC-MSC-Exos morphology. Scale bars = 100 nm and 50 nm, respectively. (E) Detection of CD9, CD63, and CD81 incorporation into hiPSC-MSC-Exos by western blotting.

Article Snippet: The effects of exosomes on fibroblasts migration were evaluated in cell responses level by western blot measurement of fibronectin (Santa Cruz Biotechnology) protein levels at 24 h. The total cellular proteins were first extracted and the cell lysates were cleared by centrifugation at 4°C and 12,000 rpm for 15 min.

Techniques: Derivative Assay, Light Microscopy, Staining, Quantitative RT-PCR, Transmission Assay, Microscopy, Western Blot

Journal: Journal of Translational Medicine

Article Title: Exosomes released from human induced pluripotent stem cells-derived MSCs facilitate cutaneous wound healing by promoting collagen synthesis and angiogenesis

doi: 10.1186/s12967-015-0417-0

Figure Lengend Snippet: The effects of exosomes on the proliferation, migration, and collagen, elastin secretion of human fibroblasts. The light microscopy images (A) and migration rates (B) of human fibroblasts into scratch sites following growth in MesenGro hMSC medium containing 0, 50, or 100 μg/mL hiPSC-MSC-Exos for 12 or 24 h. Scale bar = 250 μm. (C) Fibronectin protein expression of human fibroblasts treated with MesenGro hMSC medium containing 0, 50, or 100 μg/mL hiPSC-MSC-Exos for 24 h. (D) Human fibroblasts proliferation after growth in MesenGro hMSC medium containing 0, 50, or 100 μg/mL hiPSC-MSC-Exos was detected with a CCK-8 kit over 5 days. Secretion of Col I (E) , III (F) and elastin (G) by human fibroblasts after growth in MesenGro hMSC medium containing 0, 50, or 100 μg/mL hiPSC-MSC-Exos over 3 days. (G) The Col I (H) , III (I) and elastin (J) mRNA expression of human fibroblasts treated with MesenGro hMSC medium containing 0, 50, or 100 μg/mL hiPSC-MSC-Exos over 3 days. *P < 0.05.

Article Snippet: The effects of exosomes on fibroblasts migration were evaluated in cell responses level by western blot measurement of fibronectin (Santa Cruz Biotechnology) protein levels at 24 h. The total cellular proteins were first extracted and the cell lysates were cleared by centrifugation at 4°C and 12,000 rpm for 15 min.

Techniques: Migration, Light Microscopy, Expressing, CCK-8 Assay