Hypoxia-Induced Changes in the Bioactivity of Cytotrophoblast-Derived Exosomes (original) (raw)
Figure 1
Detection and characterization of Cytotrophoblast cell-derived exosomes.
Cytotrophoblast cells were isolated from chorionic villi obtained from first trimester pregnancies and cultured under different oxygen tension (see Methods). Exosomes were isolated from CTs culture media and characterized morphologically and using specific marker for exosome proteins. (A) Representative Western blot for exosome markers: CD63, CD9, CD81 and PLAP. Sample loading was normalized by protein content. Fractions 1 to 10, represent fractions collected after buoyant density centrifugation. (B) Protein profile of exosomal proteins and cytotrophoblast cells proteins. Exosomal proteins derived from fraction 5 to 8 (positive for exosomal marker) and cellular proteins of trophoblast cells (cells) were separated by SDS-PAGE and stained with SimplyBlueTM SafeStain. (C) Electron micrograph of exosomes isolated by ultracentrifuge and purified with a sucrose gradient (pooled exosomal pellet) from cytotrophoblast cells. In B, Scale bar 100 nm. In A, B and C, none of the experiments performed were significantly different using different oxygen tension.