{"id":{"repo_id":"helsinki","oai_identifier":"oai:helda.helsinki.fi:10138/230824"},"canonical_url":"https://search.dev.ndltd.org/etd/helsinki/oai:helda.helsinki.fi:10138/230824","repository":{"repo_id":"helsinki","name":"University of Helsinki","base_url":"https://helda.helsinki.fi/server/oai/request"},"display":{"title":"Impact of human blood plasma-derived protein corona on extracellular vesicle uptake","abstract":"Cells release different types of phospholipid bilayer-limited vesicles into the extracellular space. These are commonly referred to as extracellular vesicles (EVs). Exosomes (EXOs), ca 50-100 nm in diameter and microvesicles (MVs), ca 100-1000 nm in diameter, having different intracellular origin, are the two main subpopulations of EVs. EVs have been demonstrated to carry a range of proteins and nucleic acids subsequently delivered to recipient cells, making them attractive as drug delivery vehicles. Several mechanisms for the cellular uptake of EVs have been established. When a nanoparticle is introduced into blood plasma, plasma proteins are adsorbed to its surface, forming a protein corona. The formation of the corona is a dynamic process, governed by individual protein concentrations as well as their respective affinities for the surface. Proteins of the corona interact with surrounding cells, thus being able to influence the cellular uptake of the nanoparticle. In the current study, the uptake of PC-3-derived EVs into PC-3 cells was investigated. Moreover, the impact of a human blood plasma-derived protein corona on said uptake was assessed. EVs were isolated from collected PC-3 cell culture medium using differential centrifugation. Experiments were performed separately for MVs (20000xg EV-fraction) and EXOs (110000xg EVfraction). SDS-PAGE analysis revealed adsorption of plasma proteins to EVs, following their exposure to plasma. Prior to uptake experiments DiO-labelled EVs were either incubated or not incubated in plasma. Plasma incubation lasted overnight. PC-3 cells were then treated with either of the two EV-preparations. Following incubation, EV uptake was assessed using confocal microscopy by determining the percentage of positive fluorescent cells in cell cultures. Pre-study plasma incubation resulted in a reduced or unchanged uptake of MVs and in a reduced uptake of EXOs, when compared to their native counterparts. In conclusion, the plasma-derived protein corona was shown not to improve EV uptake. It is worth noting that the current study limits itself to the use of PC-3-derived EVs and PC-3 cells as recipient cells in uptake experiments.","abstract_html":"Cells release different types of phospholipid bilayer-limited vesicles into the extracellular space. These are commonly referred to as extracellular vesicles (EVs). Exosomes (EXOs), ca 50-100 nm in diameter and microvesicles (MVs), ca 100-1000 nm in diameter, having different intracellular origin, are the two main subpopulations of EVs. EVs have been demonstrated to carry a range of proteins and nucleic acids subsequently delivered to recipient cells, making them attractive as drug delivery vehicles. Several mechanisms for the cellular uptake of EVs have been established. When a nanoparticle is introduced into blood plasma, plasma proteins are adsorbed to its surface, forming a protein corona. The formation of the corona is a dynamic process, governed by individual protein concentrations as well as their respective affinities for the surface. Proteins of the corona interact with surrounding cells, thus being able to influence the cellular uptake of the nanoparticle. In the current study, the uptake of PC-3-derived EVs into PC-3 cells was investigated. Moreover, the impact of a human blood plasma-derived protein corona on said uptake was assessed. EVs were isolated from collected PC-3 cell culture medium using differential centrifugation. Experiments were performed separately for MVs (20000xg EV-fraction) and EXOs (110000xg EVfraction). SDS-PAGE analysis revealed adsorption of plasma proteins to EVs, following their exposure to plasma. Prior to uptake experiments DiO-labelled EVs were either incubated or not incubated in plasma. Plasma incubation lasted overnight. PC-3 cells were then treated with either of the two EV-preparations. Following incubation, EV uptake was assessed using confocal microscopy by determining the percentage of positive fluorescent cells in cell cultures. Pre-study plasma incubation resulted in a reduced or unchanged uptake of MVs and in a reduced uptake of EXOs, when compared to their native counterparts. In conclusion, the plasma-derived protein corona was shown not to improve EV uptake. It is worth noting that the current study limits itself to the use of PC-3-derived EVs and PC-3 cells as recipient cells in uptake experiments.","abstract_has_math":false,"creators":["Somersalo, Petter"],"institution":"Helsingfors universitet","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Helsingin yliopisto, Farmasian tiedekunta","University of Helsinki, Faculty of Pharmacy","Helsingfors universitet, Farmaceutiska fakulteten"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2017,"date_issued":"2017","date_published":"2017","updated_at":"2026-07-27T19:56:06Z","subjects":["extracellular vesicles","cellular uptake","protein adsorption","protein corona","blood plasma","solunulkoiset vesikkelit","soluunotto","proteiiniadsorptio","proteiinikorona","veriplasma"],"languages":["eng"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["URN:NBN:fi-fe201801151392"],"render_values":[{"text":"URN:NBN:fi-fe201801151392","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10138/230824","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Helsingin yliopisto, Farmasian tiedekunta","University of Helsinki, Faculty of Pharmacy","Helsingfors universitet, Farmaceutiska fakulteten"]},{"key":"dc:creator","label":"Author","values":["Somersalo, Petter"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2017"]},{"key":"dc:publisher","label":"Institution","values":["Helsingfors universitet","University of Helsinki","Helsingin yliopisto"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["extracellular vesicles","cellular uptake","protein adsorption","protein corona","blood plasma","solunulkoiset vesikkelit","soluunotto","proteiiniadsorptio","proteiinikorona","veriplasma"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["URN:NBN:fi-fe201801151392","http://hdl.handle.net/10138/230824"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Cells release different types of phospholipid bilayer-limited vesicles into the extracellular space. These are commonly referred to as extracellular vesicles (EVs). Exosomes (EXOs), ca 50-100 nm in diameter and microvesicles (MVs), ca 100-1000 nm in diameter, having different intracellular origin, are the two main subpopulations of EVs. EVs have been demonstrated to carry a range of proteins and nucleic acids subsequently delivered to recipient cells, making them attractive as drug delivery vehicles. Several mechanisms for the cellular uptake of EVs have been established. When a nanoparticle is introduced into blood plasma, plasma proteins are adsorbed to its surface, forming a protein corona. The formation of the corona is a dynamic process, governed by individual protein concentrations as well as their respective affinities for the surface. Proteins of the corona interact with surrounding cells, thus being able to influence the cellular uptake of the nanoparticle. In the current study, the uptake of PC-3-derived EVs into PC-3 cells was investigated. Moreover, the impact of a human blood plasma-derived protein corona on said uptake was assessed. EVs were isolated from collected PC-3 cell culture medium using differential centrifugation. Experiments were performed separately for MVs (20000xg EV-fraction) and EXOs (110000xg EVfraction). SDS-PAGE analysis revealed adsorption of plasma proteins to EVs, following their exposure to plasma. Prior to uptake experiments DiO-labelled EVs were either incubated or not incubated in plasma. Plasma incubation lasted overnight. PC-3 cells were then treated with either of the two EV-preparations. Following incubation, EV uptake was assessed using confocal microscopy by determining the percentage of positive fluorescent cells in cell cultures. Pre-study plasma incubation resulted in a reduced or unchanged uptake of MVs and in a reduced uptake of EXOs, when compared to their native counterparts. In conclusion, the plasma-derived protein corona was shown not to improve EV uptake. It is worth noting that the current study limits itself to the use of PC-3-derived EVs and PC-3 cells as recipient cells in uptake experiments.","Solunulkoiset vesikkelit (extracellular vesicles, EVs) ovat solujen tuottamia, solunulkoiseen tilaan vapautuvia vesikkeleitä. Syntymekanisminsa perusteella EV:t voidaan jakaa useisiin eri tyyppeihin, mukaan lukien eksosomit ja mikrovesikkelit. Halkaisijaltaan eksosomit ovat n. 50-100 nm, ja mikrovesikkelit n. 100-1000 nm. EV:iden on todettu kuljettavan eri proteiineja ja nukleiinihappoja solusta toiseen, mikä tekeekin niiden hyödyntämisestä lääkkeenkantajina houkuttelevaa. Nanopartikkelin joutuessa kosketuksiin veren plasman kanssa sen pintaan adsorboituu plasman proteiineja, jotka muodostavat proteiinikoronan nanopartikkelin ympärille. Proteiinikoronan muodostuminen on dynaaminen prosessi, johon vaikuttavat yksittäisten proteiinien pitoisuus ja affiniteetti pintaa kohtaan. Nanopartikkelin pintaan adsorboituneet proteiinit vuorovaikuttavat ympäröivien solujen kanssa ja näin ollen vaikuttavat nanopartikkelin soluunottoon. Työssä tutkittiin PC-3-solujen EV:iden soluunottoa samaisessa solulinjassa. Erityisesti, ihmisen veriplasmasta lähtöisin olevan proteiinikoronan vaikutusta kyseiseen soluunottoon arvioitiin. EV:t eristettiin kerätystä PC-3-solujen kasvatusmediumista differentiaalisentrifugaatiota käyttäen. Kokeet suoritettiin erikseen käyttäen mikrovesikkeleitä (20000xg EV-fraktio) ja eksosomeja (110000xg EV-fraktio). SDS-PAGE-analyysin avulla todennettiin plasmaproteiinien adsorptio EV:ihin niiden plasmassa inkuboinnin seurauksena. Soluunottokokeissa käytettiin DiO:lla leimattuja EV:itä, joita oli joko inkuboitu tai ei inkuboitu plasmassa. PC-3-solut altistettiin jommallekummalle kyseisistä EVpreparaateista. EV:iden soluunotto määritettiin konfokaalimikroskopiaa käyttäen laskemalla fluoresenssin suhteen positiivisten solujen osuus soluviljelmässä. Soluunottokokeita edeltävän plasmainkuboinnin vaikutus EV:iden soluunottoon oli negatiivinen tai olematon mikrovesikkelien tapauksessa ja negatiivinen eksosomien tapauksessa. Toisin sanoen veriplasmasta lähtöisin olevan proteiinikoronan ei todettu parantavan EV:iden soluunottoa. On kuitenkin syytä huomioida kyseisten soluunottokokeiden tulosten rajoittuvan PC-3-soluihin, ja PC-3-solujen EV:iden käyttöön."]},{"key":"dc:title","label":"Title","values":["Impact of human blood plasma-derived protein corona on extracellular vesicle uptake"]}]}],"canonical_facts":{"dc:contributor":["Helsingin yliopisto, Farmasian tiedekunta","University of Helsinki, Faculty of Pharmacy","Helsingfors universitet, Farmaceutiska fakulteten"],"dc:creator":["Somersalo, Petter"],"dc:date.issued":["2017"],"dc:description.abstract":["Cells release different types of phospholipid bilayer-limited vesicles into the extracellular space. These are commonly referred to as extracellular vesicles (EVs). Exosomes (EXOs), ca 50-100 nm in diameter and microvesicles (MVs), ca 100-1000 nm in diameter, having different intracellular origin, are the two main subpopulations of EVs. EVs have been demonstrated to carry a range of proteins and nucleic acids subsequently delivered to recipient cells, making them attractive as drug delivery vehicles. Several mechanisms for the cellular uptake of EVs have been established. When a nanoparticle is introduced into blood plasma, plasma proteins are adsorbed to its surface, forming a protein corona. The formation of the corona is a dynamic process, governed by individual protein concentrations as well as their respective affinities for the surface. Proteins of the corona interact with surrounding cells, thus being able to influence the cellular uptake of the nanoparticle. In the current study, the uptake of PC-3-derived EVs into PC-3 cells was investigated. Moreover, the impact of a human blood plasma-derived protein corona on said uptake was assessed. EVs were isolated from collected PC-3 cell culture medium using differential centrifugation. Experiments were performed separately for MVs (20000xg EV-fraction) and EXOs (110000xg EVfraction). SDS-PAGE analysis revealed adsorption of plasma proteins to EVs, following their exposure to plasma. Prior to uptake experiments DiO-labelled EVs were either incubated or not incubated in plasma. Plasma incubation lasted overnight. PC-3 cells were then treated with either of the two EV-preparations. Following incubation, EV uptake was assessed using confocal microscopy by determining the percentage of positive fluorescent cells in cell cultures. Pre-study plasma incubation resulted in a reduced or unchanged uptake of MVs and in a reduced uptake of EXOs, when compared to their native counterparts. In conclusion, the plasma-derived protein corona was shown not to improve EV uptake. It is worth noting that the current study limits itself to the use of PC-3-derived EVs and PC-3 cells as recipient cells in uptake experiments.","Solunulkoiset vesikkelit (extracellular vesicles, EVs) ovat solujen tuottamia, solunulkoiseen tilaan vapautuvia vesikkeleitä. Syntymekanisminsa perusteella EV:t voidaan jakaa useisiin eri tyyppeihin, mukaan lukien eksosomit ja mikrovesikkelit. Halkaisijaltaan eksosomit ovat n. 50-100 nm, ja mikrovesikkelit n. 100-1000 nm. EV:iden on todettu kuljettavan eri proteiineja ja nukleiinihappoja solusta toiseen, mikä tekeekin niiden hyödyntämisestä lääkkeenkantajina houkuttelevaa. Nanopartikkelin joutuessa kosketuksiin veren plasman kanssa sen pintaan adsorboituu plasman proteiineja, jotka muodostavat proteiinikoronan nanopartikkelin ympärille. Proteiinikoronan muodostuminen on dynaaminen prosessi, johon vaikuttavat yksittäisten proteiinien pitoisuus ja affiniteetti pintaa kohtaan. Nanopartikkelin pintaan adsorboituneet proteiinit vuorovaikuttavat ympäröivien solujen kanssa ja näin ollen vaikuttavat nanopartikkelin soluunottoon. Työssä tutkittiin PC-3-solujen EV:iden soluunottoa samaisessa solulinjassa. Erityisesti, ihmisen veriplasmasta lähtöisin olevan proteiinikoronan vaikutusta kyseiseen soluunottoon arvioitiin. EV:t eristettiin kerätystä PC-3-solujen kasvatusmediumista differentiaalisentrifugaatiota käyttäen. Kokeet suoritettiin erikseen käyttäen mikrovesikkeleitä (20000xg EV-fraktio) ja eksosomeja (110000xg EV-fraktio). SDS-PAGE-analyysin avulla todennettiin plasmaproteiinien adsorptio EV:ihin niiden plasmassa inkuboinnin seurauksena. Soluunottokokeissa käytettiin DiO:lla leimattuja EV:itä, joita oli joko inkuboitu tai ei inkuboitu plasmassa. PC-3-solut altistettiin jommallekummalle kyseisistä EVpreparaateista. EV:iden soluunotto määritettiin konfokaalimikroskopiaa käyttäen laskemalla fluoresenssin suhteen positiivisten solujen osuus soluviljelmässä. Soluunottokokeita edeltävän plasmainkuboinnin vaikutus EV:iden soluunottoon oli negatiivinen tai olematon mikrovesikkelien tapauksessa ja negatiivinen eksosomien tapauksessa. Toisin sanoen veriplasmasta lähtöisin olevan proteiinikoronan ei todettu parantavan EV:iden soluunottoa. On kuitenkin syytä huomioida kyseisten soluunottokokeiden tulosten rajoittuvan PC-3-soluihin, ja PC-3-solujen EV:iden käyttöön."],"dc:identifier.uri":["URN:NBN:fi-fe201801151392","http://hdl.handle.net/10138/230824"],"dc:language.iso":["eng"],"dc:publisher":["Helsingfors universitet","University of Helsinki","Helsingin yliopisto"],"dc:subject":["extracellular vesicles","cellular uptake","protein adsorption","protein corona","blood plasma","solunulkoiset vesikkelit","soluunotto","proteiiniadsorptio","proteiinikorona","veriplasma"],"dc:title":["Impact of human blood plasma-derived protein corona on extracellular vesicle uptake"]},"updated_at":"2026-07-27T19:56:06Z"}