From bench to bedside: a systematic approach to increased laboratory exosome production
Background: Over the last years, interest for microvesicles and exosomes has significantly increased as they revealed a high therapeutical potential for several clinical conditions, such as haemorrhagic shock, cancer, among others. The bottleneck for preclinical and clinical testing remains the reli...
Gespeichert in:
| Veröffentlicht in: | Journal of extracellular vesicles 2018-01, Vol.7, p.224-224 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 224 |
|---|---|
| container_issue | |
| container_start_page | 224 |
| container_title | Journal of extracellular vesicles |
| container_volume | 7 |
| creator | Schuh, Christina M A P Tapia, Rafael Khoury, Maroun |
| description | Background: Over the last years, interest for microvesicles and exosomes has significantly increased as they revealed a high therapeutical potential for several clinical conditions, such as haemorrhagic shock, cancer, among others. The bottleneck for preclinical and clinical testing remains the reliable production of exosomes with consistent quality, as existing processes not only are unreliable concerning purity and scaling ( |
| format | Article |
| fullrecord | <record><control><sourceid>proquest</sourceid><recordid>TN_cdi_proquest_journals_2116616672</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2116616672</sourcerecordid><originalsourceid>FETCH-proquest_journals_21166166723</originalsourceid><addsrcrecordid>eNqNyk0KwjAQBeAgCBbtHQKuC02r_XErFg8guCxpMmJKm6mZFOztDegBHB7Mg_etWJSlqUjytKw2LCbq03D1QRyrOmL3xuHIO7DqyT2GosloOHHJaSEPo_RGcTlNDuVXGKscSALNB9mhkx7dwuGNhCPwwPSsvEG7Y-uHHAji39-yfXO5na9JIK8ZyLc9zs6Gqc2EKIqQMsv_Ux94g0IX</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2116616672</pqid></control><display><type>article</type><title>From bench to bedside: a systematic approach to increased laboratory exosome production</title><source>Taylor & Francis Open Access</source><source>DOAJ Directory of Open Access Journals</source><source>Co-Action Open Access Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Wiley-Blackwell Open Access Titles</source><source>Wiley Online Library All Journals</source><source>PubMed Central</source><creator>Schuh, Christina M A P ; Tapia, Rafael ; Khoury, Maroun</creator><creatorcontrib>Schuh, Christina M A P ; Tapia, Rafael ; Khoury, Maroun</creatorcontrib><description>Background: Over the last years, interest for microvesicles and exosomes has significantly increased as they revealed a high therapeutical potential for several clinical conditions, such as haemorrhagic shock, cancer, among others. The bottleneck for preclinical and clinical testing remains the reliable production of exosomes with consistent quality, as existing processes not only are unreliable concerning purity and scaling (<500 ml), but also are unreproducible due to batch-differences. The aim of our study was to design a process and evaluation system for optimized laboratory scale production of exosomes that can be transferred to a GMP environment. Methods: Mesenchymal stem cells derived from menstrual fluid were cultivated under classic cell culture conditions or using microcarrier support, chosen under the prerequisite to be transferrable into GMP: BioNoc, Cytodex 3 and Capex. Culture conditions were evaluated assessing the exosome yield (NanoSight), exosome composition (Western blot), as well as cell viability (MTT assay) and onset of cell senescence (X-Gal assay). Ultracentrifugation of supernatants and its variations (gradient centrifugations, centricon prepurification) is the most abundantly used method for exosome isolation. Tangential flow filtration represents a GMP-compliable alternative to purify exosomes from small (500 ml) to large (101) volumes and through defined kDa cut-offs-modulate the composition. Following purification, exosomes can be stored in native or lyophilized state. Results: We will present results on how microcarrier implementation improves exosome yield and cell viability, as well as data on tangential flow filtration compared to ultracentrifugation. Summary/Conclusion: Our process offers a systematic approach to step-by step optimize exosome production regarding yield and purity, and-due to its GMP-compliable techniques - facilitating the translation of exosome therapies into the clinics.</description><identifier>EISSN: 2001-3078</identifier><language>eng</language><publisher>Abingdon: John Wiley & Sons, Inc</publisher><subject>Cancer ; Cell culture ; Exosomes ; Filtration ; Laboratories ; Menstruation ; Mesenchyme ; Purification ; Scaling ; Senescence ; Stem cells ; Ultracentrifugation</subject><ispartof>Journal of extracellular vesicles, 2018-01, Vol.7, p.224-224</ispartof><rights>Copyright Taylor & Francis Ltd. 2018</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids></links><search><creatorcontrib>Schuh, Christina M A P</creatorcontrib><creatorcontrib>Tapia, Rafael</creatorcontrib><creatorcontrib>Khoury, Maroun</creatorcontrib><title>From bench to bedside: a systematic approach to increased laboratory exosome production</title><title>Journal of extracellular vesicles</title><description>Background: Over the last years, interest for microvesicles and exosomes has significantly increased as they revealed a high therapeutical potential for several clinical conditions, such as haemorrhagic shock, cancer, among others. The bottleneck for preclinical and clinical testing remains the reliable production of exosomes with consistent quality, as existing processes not only are unreliable concerning purity and scaling (<500 ml), but also are unreproducible due to batch-differences. The aim of our study was to design a process and evaluation system for optimized laboratory scale production of exosomes that can be transferred to a GMP environment. Methods: Mesenchymal stem cells derived from menstrual fluid were cultivated under classic cell culture conditions or using microcarrier support, chosen under the prerequisite to be transferrable into GMP: BioNoc, Cytodex 3 and Capex. Culture conditions were evaluated assessing the exosome yield (NanoSight), exosome composition (Western blot), as well as cell viability (MTT assay) and onset of cell senescence (X-Gal assay). Ultracentrifugation of supernatants and its variations (gradient centrifugations, centricon prepurification) is the most abundantly used method for exosome isolation. Tangential flow filtration represents a GMP-compliable alternative to purify exosomes from small (500 ml) to large (101) volumes and through defined kDa cut-offs-modulate the composition. Following purification, exosomes can be stored in native or lyophilized state. Results: We will present results on how microcarrier implementation improves exosome yield and cell viability, as well as data on tangential flow filtration compared to ultracentrifugation. Summary/Conclusion: Our process offers a systematic approach to step-by step optimize exosome production regarding yield and purity, and-due to its GMP-compliable techniques - facilitating the translation of exosome therapies into the clinics.</description><subject>Cancer</subject><subject>Cell culture</subject><subject>Exosomes</subject><subject>Filtration</subject><subject>Laboratories</subject><subject>Menstruation</subject><subject>Mesenchyme</subject><subject>Purification</subject><subject>Scaling</subject><subject>Senescence</subject><subject>Stem cells</subject><subject>Ultracentrifugation</subject><issn>2001-3078</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNyk0KwjAQBeAgCBbtHQKuC02r_XErFg8guCxpMmJKm6mZFOztDegBHB7Mg_etWJSlqUjytKw2LCbq03D1QRyrOmL3xuHIO7DqyT2GosloOHHJaSEPo_RGcTlNDuVXGKscSALNB9mhkx7dwuGNhCPwwPSsvEG7Y-uHHAji39-yfXO5na9JIK8ZyLc9zs6Gqc2EKIqQMsv_Ux94g0IX</recordid><startdate>20180101</startdate><enddate>20180101</enddate><creator>Schuh, Christina M A P</creator><creator>Tapia, Rafael</creator><creator>Khoury, Maroun</creator><general>John Wiley & Sons, Inc</general><scope>7QP</scope><scope>8FE</scope><scope>8FH</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>LK8</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope></search><sort><creationdate>20180101</creationdate><title>From bench to bedside: a systematic approach to increased laboratory exosome production</title><author>Schuh, Christina M A P ; Tapia, Rafael ; Khoury, Maroun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-proquest_journals_21166166723</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Cancer</topic><topic>Cell culture</topic><topic>Exosomes</topic><topic>Filtration</topic><topic>Laboratories</topic><topic>Menstruation</topic><topic>Mesenchyme</topic><topic>Purification</topic><topic>Scaling</topic><topic>Senescence</topic><topic>Stem cells</topic><topic>Ultracentrifugation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schuh, Christina M A P</creatorcontrib><creatorcontrib>Tapia, Rafael</creatorcontrib><creatorcontrib>Khoury, Maroun</creatorcontrib><collection>Calcium & Calcified Tissue Abstracts</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Biological Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><jtitle>Journal of extracellular vesicles</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schuh, Christina M A P</au><au>Tapia, Rafael</au><au>Khoury, Maroun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>From bench to bedside: a systematic approach to increased laboratory exosome production</atitle><jtitle>Journal of extracellular vesicles</jtitle><date>2018-01-01</date><risdate>2018</risdate><volume>7</volume><spage>224</spage><epage>224</epage><pages>224-224</pages><eissn>2001-3078</eissn><abstract>Background: Over the last years, interest for microvesicles and exosomes has significantly increased as they revealed a high therapeutical potential for several clinical conditions, such as haemorrhagic shock, cancer, among others. The bottleneck for preclinical and clinical testing remains the reliable production of exosomes with consistent quality, as existing processes not only are unreliable concerning purity and scaling (<500 ml), but also are unreproducible due to batch-differences. The aim of our study was to design a process and evaluation system for optimized laboratory scale production of exosomes that can be transferred to a GMP environment. Methods: Mesenchymal stem cells derived from menstrual fluid were cultivated under classic cell culture conditions or using microcarrier support, chosen under the prerequisite to be transferrable into GMP: BioNoc, Cytodex 3 and Capex. Culture conditions were evaluated assessing the exosome yield (NanoSight), exosome composition (Western blot), as well as cell viability (MTT assay) and onset of cell senescence (X-Gal assay). Ultracentrifugation of supernatants and its variations (gradient centrifugations, centricon prepurification) is the most abundantly used method for exosome isolation. Tangential flow filtration represents a GMP-compliable alternative to purify exosomes from small (500 ml) to large (101) volumes and through defined kDa cut-offs-modulate the composition. Following purification, exosomes can be stored in native or lyophilized state. Results: We will present results on how microcarrier implementation improves exosome yield and cell viability, as well as data on tangential flow filtration compared to ultracentrifugation. Summary/Conclusion: Our process offers a systematic approach to step-by step optimize exosome production regarding yield and purity, and-due to its GMP-compliable techniques - facilitating the translation of exosome therapies into the clinics.</abstract><cop>Abingdon</cop><pub>John Wiley & Sons, Inc</pub></addata></record> |
| fulltext | fulltext |
| identifier | EISSN: 2001-3078 |
| ispartof | Journal of extracellular vesicles, 2018-01, Vol.7, p.224-224 |
| issn | 2001-3078 |
| language | eng |
| recordid | cdi_proquest_journals_2116616672 |
| source | Taylor & Francis Open Access; DOAJ Directory of Open Access Journals; Co-Action Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Wiley-Blackwell Open Access Titles; Wiley Online Library All Journals; PubMed Central |
| subjects | Cancer Cell culture Exosomes Filtration Laboratories Menstruation Mesenchyme Purification Scaling Senescence Stem cells Ultracentrifugation |
| title | From bench to bedside: a systematic approach to increased laboratory exosome production |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-05T17%3A14%3A18IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=From%20bench%20to%20bedside:%20a%20systematic%20approach%20to%20increased%20laboratory%20exosome%20production&rft.jtitle=Journal%20of%20extracellular%20vesicles&rft.au=Schuh,%20Christina%20M%20A%20P&rft.date=2018-01-01&rft.volume=7&rft.spage=224&rft.epage=224&rft.pages=224-224&rft.eissn=2001-3078&rft_id=info:doi/&rft_dat=%3Cproquest%3E2116616672%3C/proquest%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2116616672&rft_id=info:pmid/&rfr_iscdi=true |