Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine
Phosphatidylserine (PS) has skewed distributions in the plasma membrane and is preferentially located in the inner leaflet of normal cells. Tumour cells, however, expose PS at the outer leaflet of cell surfaces, thereby potentially modulating the bio-signalling of cells. Interestingly, exosomes – or...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2019-12-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/20013078.2019.1579541 |
| _version_ | 1818907297350418432 |
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| author | Sachiko Matsumura Tamiko Minamisawa Kanako Suga Hiromi Kishita Takanori Akagi Takanori Ichiki Yuki Ichikawa Kiyotaka Shiba |
| author_facet | Sachiko Matsumura Tamiko Minamisawa Kanako Suga Hiromi Kishita Takanori Akagi Takanori Ichiki Yuki Ichikawa Kiyotaka Shiba |
| author_sort | Sachiko Matsumura |
| collection | DOAJ |
| description | Phosphatidylserine (PS) has skewed distributions in the plasma membrane and is preferentially located in the inner leaflet of normal cells. Tumour cells, however, expose PS at the outer leaflet of cell surfaces, thereby potentially modulating the bio-signalling of cells. Interestingly, exosomes – or, more properly, small extracellular vesicles (sEVs) – which are secreted from tumour cells, are enriched with externalized PS, have been proposed as being involved in the progression of cancers, and could be used as a marker for tumour diagnostics. However, the sEV fractions prepared from various methods are composed of different subtypes of vesicles, and knowledge about the subtypes enriched with exposed PS is still limited. Here, we differentiated sEVs from cancer cell lines by density gradient centrifugation and characterized the separated fractions by using gold-labelling of PS in atomic force microscopy, thrombin generation assay, size and zeta potential measurements, and western blot analysis. These analyses revealed a previously unreported PS+-enriched sEV subtype, which is characterized by a lower density than that of canonical exosomes (1.06 g/ml vs. 1.08 g/ml), larger size (122 nm vs. 105 nm), more negative zeta potential (−28 mV vs. −21 mV), and lower abundance of canonical exosomal markers. The identification of the PS-exposed subtype of sEVs will provide deeper insight into the role of EVs in tumour biology and enhance the development of EV-based tumour diagnosis and therapy. |
| first_indexed | 2024-12-19T21:52:53Z |
| format | Article |
| id | doaj.art-10f0bee81c0e4ec0ac010788a599f95d |
| institution | Directory Open Access Journal |
| issn | 2001-3078 |
| language | English |
| last_indexed | 2024-12-19T21:52:53Z |
| publishDate | 2019-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj.art-10f0bee81c0e4ec0ac010788a599f95d2022-12-21T20:04:21ZengWileyJournal of Extracellular Vesicles2001-30782019-12-018110.1080/20013078.2019.15795411579541Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserineSachiko Matsumura0Tamiko Minamisawa1Kanako Suga2Hiromi Kishita3Takanori Akagi4Takanori Ichiki5Yuki Ichikawa6Kiyotaka Shiba7Cancer Institute, Japanese Foundation for Cancer ResearchCancer Institute, Japanese Foundation for Cancer ResearchCancer Institute, Japanese Foundation for Cancer ResearchThe University of TokyoThe University of TokyoThe University of TokyoIMRA America, IncCancer Institute, Japanese Foundation for Cancer ResearchPhosphatidylserine (PS) has skewed distributions in the plasma membrane and is preferentially located in the inner leaflet of normal cells. Tumour cells, however, expose PS at the outer leaflet of cell surfaces, thereby potentially modulating the bio-signalling of cells. Interestingly, exosomes – or, more properly, small extracellular vesicles (sEVs) – which are secreted from tumour cells, are enriched with externalized PS, have been proposed as being involved in the progression of cancers, and could be used as a marker for tumour diagnostics. However, the sEV fractions prepared from various methods are composed of different subtypes of vesicles, and knowledge about the subtypes enriched with exposed PS is still limited. Here, we differentiated sEVs from cancer cell lines by density gradient centrifugation and characterized the separated fractions by using gold-labelling of PS in atomic force microscopy, thrombin generation assay, size and zeta potential measurements, and western blot analysis. These analyses revealed a previously unreported PS+-enriched sEV subtype, which is characterized by a lower density than that of canonical exosomes (1.06 g/ml vs. 1.08 g/ml), larger size (122 nm vs. 105 nm), more negative zeta potential (−28 mV vs. −21 mV), and lower abundance of canonical exosomal markers. The identification of the PS-exposed subtype of sEVs will provide deeper insight into the role of EVs in tumour biology and enhance the development of EV-based tumour diagnosis and therapy.http://dx.doi.org/10.1080/20013078.2019.1579541atomic force microscopydensity gradient centrifugationexosomesextracellular vesiclesisolationphosphatidylserinethrombin generation |
| spellingShingle | Sachiko Matsumura Tamiko Minamisawa Kanako Suga Hiromi Kishita Takanori Akagi Takanori Ichiki Yuki Ichikawa Kiyotaka Shiba Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine Journal of Extracellular Vesicles atomic force microscopy density gradient centrifugation exosomes extracellular vesicles isolation phosphatidylserine thrombin generation |
| title | Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine |
| title_full | Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine |
| title_fullStr | Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine |
| title_full_unstemmed | Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine |
| title_short | Subtypes of tumour cell-derived small extracellular vesicles having differently externalized phosphatidylserine |
| title_sort | subtypes of tumour cell derived small extracellular vesicles having differently externalized phosphatidylserine |
| topic | atomic force microscopy density gradient centrifugation exosomes extracellular vesicles isolation phosphatidylserine thrombin generation |
| url | http://dx.doi.org/10.1080/20013078.2019.1579541 |
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