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PMID:21276792

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Citation

Atay, S, Gercel-Taylor, C, Kesimer, M and Taylor, DD (2011) Morphologic and proteomic characterization of exosomes released by cultured extravillous trophoblast cells. Exp. Cell Res. 317:1192-202

Abstract

Exosomes represent an important intercellular communication vehicle, mediating events essential for the decidual microenvironment. While we have demonstrated exosome induction of pro-inflammatory cytokines, to date, no extensive characterization of trophoblast-derived exosomes has been provided. Our objective was to provide a morphologic and proteomic characterization of these exosomes. Exosomes were isolated from the conditioned media of Swan71 human trophoblast cells by ultrafiltration and ultracentrifugation. These were analyzed for density (sucrose density gradient centrifugation), morphology (electron microscopy), size (dynamic light scattering) and protein composition (Ion Trap mass spectrometry and western immunoblotting). Based on density gradient centrifugation, microvesicles from Sw71 cells exhibit a density between 1.134 and 1.173g/ml. Electron microscopy demonstrated that microvesicles from Sw71 cells exhibit the characteristic cup-shaped morphology of exosomes. Dynamic light scattering showed a bell-shaped curve, indicating a homogeneous population with a mean size of 165nm ± 0.5nm. Ion Trap mass spectrometry demonstrated the presence of exosome marker proteins (including CD81, Alix, cytoskeleton related proteins, and Rab family). The MS results were confirmed by western immunoblotting. Based on morphology, density, size and protein composition, we defined the release of exosomes from extravillous trophoblast cells and provide their first extensive characterization. This characterization is essential in furthering our understanding of "normal" early pregnancy.

Links

PubMed Online version:10.1016/j.yexcr.2011.01.014

Keywords

Cells, Cultured; Centrifugation, Density Gradient; Exosomes/chemistry; Exosomes/ultrastructure; Female; Gene Regulatory Networks; Humans; Mass Spectrometry/methods; Particle Size; Pregnancy; Proteome/analysis; Proteomics/methods; Trophoblasts/cytology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:FINC

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:ALBU

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:PAI1

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:ICAM1

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:CH60

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:HSP74

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:CD81

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:PDC6I

located_in

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:CH60

part_of

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:FINC

part_of

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:ALBU

part_of

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000351)

Seeded From UniProt

complete

HUMAN:HSP74

part_of

GO:0070062: extracellular exosome

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000351)

Seeded From UniProt

complete

Notes

See also

References

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