Cytoplasmic expression of EGFP in dendritic cells transfected with in vitro transcribed mRNA or cellular total RNA extracted from EGFP expressing leukemia cells

The present study was designed for identifying the protein synthesis in cytoplasm of dendritic cells transfected with in vitro transcribed mRNA and cellular total RNA extracted from tumor cells. Dendritic cells were generated from cord blood-CD34+ cells by culture with GM-CSF, SCF, and TNF-alpha, or...

Full description

Saved in:
Bibliographic Details
Published in:Medical oncology (Northwood, London, England) London, England), 2003-01, Vol.20 (4), p.335-348
Main Authors: Takahashi, Masuhiro, Narita, Miwako, Ayres, Flavio, Satoh, Naoko, Abe, Takashi, Yanao, Toshio, Furukawa, Tatsuo, Toba, Ken, Hirohashi, Takeshi, Aizawa, Yoshifusa
Format: Article
Language:English
Subjects:
Dendritic Cells - immunology
Dendritic Cells - metabolism
Electroporation - methods
Green Fluorescent Proteins
Humans
Immunotherapy - methods
Leukemia
Luminescent Proteins - metabolism
Oncology
RNA, Messenger - genetics
RNA, Messenger - isolation & purification
Transfection - methods
Tumor Cells, Cultured - metabolism
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c312t-d9dcfa6d98c4662599ac123e527a061ea9389c90f938a1d2c94a10fa8985b7163
cites
container_end_page 348
container_issue 4
container_start_page 335
container_title Medical oncology (Northwood, London, England)
container_volume 20
creator Takahashi, Masuhiro
Narita, Miwako
Ayres, Flavio
Satoh, Naoko
Abe, Takashi
Yanao, Toshio
Furukawa, Tatsuo
Toba, Ken
Hirohashi, Takeshi
Aizawa, Yoshifusa
description The present study was designed for identifying the protein synthesis in cytoplasm of dendritic cells transfected with in vitro transcribed mRNA and cellular total RNA extracted from tumor cells. Dendritic cells were generated from cord blood-CD34+ cells by culture with GM-CSF, SCF, and TNF-alpha, or from peripheral blood adherent cells or CD14+ cells by culture with GM-CSF and IL-4. Dendritic cells were transfected with in vitro transcribed EGFP mRNA or cellular total RNA, which was isolated from EGFP expressing K562, by electroporation using a square-wave pulse. Optimal in vitro transcribed EGFP mRNA transfection efficiency (>90%) was observed in a single electroporation of 1.75 kV/cm (electric field strength) with a pulse width of 250 micros. Although the intensity of EGFP expression in dendritic cells transfected with cellular total RNA was less compared with that in dendritic cells transfected with in vitro transcribed EGFP mRNA, a definite cytoplasmic synthesis of EGFP was demonstrated in dendritic cells transfected with cellular total RNA. The visual identification of cytoplasmic expression of cellular total RNA in dendritic cells revealed that electroporation of tumor cell-derived RNA could be a useful tool to load dendritic cells with tumor antigens for establishing an efficient dendritic cell-based tumor immunotherapy.
doi_str_mv 10.1385/MO:20:4:335
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_71502543</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2394108251</sourcerecordid><originalsourceid>FETCH-LOGICAL-c312t-d9dcfa6d98c4662599ac123e527a061ea9389c90f938a1d2c94a10fa8985b7163</originalsourceid><addsrcrecordid>eNpdkUlPwzAQhS0EYj9xRxYHLijgJU7i3qqqLBKlCIHEzXIdB1ySuNgOy7_hp-KSIiROY_l9M280D4ADjE4xLdjZZDogaJAOKGVrYBszxhNM8eN6fFOWJ4hlaAvseD9HiGBG-CbYwmmOM0T4NvgafQa7qKVvjIL6Y-G098a20FZwfHF-C00LS92WzoSoK13XHgYnW19pFXQJ3014XjJvJjjbK8qZWVSau5shtO6np6ulg8EGWcPlr_6I4E975WzT-_xat0-w1t2Lbozs7fbARiVrr_dXdRc8nI_vR5fJ9fTiajS8ThTFJCQlL1Uls5IXKs0ywjiXChOqGcklyrCWnBZccVTFKnFJFE8lRpUseMFm8Rh0Fxz3cxfOvnbaB9EYv9xAttp2XuSYIcJSGsGjf-Dcdq6Nu4kiZyzHhPAInfSQctZ7pyuxcKaR7lNgJJapiclUECRSEVOL9OFqZDdrdPnHrmKi35ZMk_E</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>875571229</pqid></control><display><type>article</type><title>Cytoplasmic expression of EGFP in dendritic cells transfected with in vitro transcribed mRNA or cellular total RNA extracted from EGFP expressing leukemia cells</title><source>Springer Link</source><creator>Takahashi, Masuhiro ; Narita, Miwako ; Ayres, Flavio ; Satoh, Naoko ; Abe, Takashi ; Yanao, Toshio ; Furukawa, Tatsuo ; Toba, Ken ; Hirohashi, Takeshi ; Aizawa, Yoshifusa</creator><creatorcontrib>Takahashi, Masuhiro ; Narita, Miwako ; Ayres, Flavio ; Satoh, Naoko ; Abe, Takashi ; Yanao, Toshio ; Furukawa, Tatsuo ; Toba, Ken ; Hirohashi, Takeshi ; Aizawa, Yoshifusa</creatorcontrib><description>The present study was designed for identifying the protein synthesis in cytoplasm of dendritic cells transfected with in vitro transcribed mRNA and cellular total RNA extracted from tumor cells. Dendritic cells were generated from cord blood-CD34+ cells by culture with GM-CSF, SCF, and TNF-alpha, or from peripheral blood adherent cells or CD14+ cells by culture with GM-CSF and IL-4. Dendritic cells were transfected with in vitro transcribed EGFP mRNA or cellular total RNA, which was isolated from EGFP expressing K562, by electroporation using a square-wave pulse. Optimal in vitro transcribed EGFP mRNA transfection efficiency (&gt;90%) was observed in a single electroporation of 1.75 kV/cm (electric field strength) with a pulse width of 250 micros. Although the intensity of EGFP expression in dendritic cells transfected with cellular total RNA was less compared with that in dendritic cells transfected with in vitro transcribed EGFP mRNA, a definite cytoplasmic synthesis of EGFP was demonstrated in dendritic cells transfected with cellular total RNA. The visual identification of cytoplasmic expression of cellular total RNA in dendritic cells revealed that electroporation of tumor cell-derived RNA could be a useful tool to load dendritic cells with tumor antigens for establishing an efficient dendritic cell-based tumor immunotherapy.</description><identifier>ISSN: 1357-0560</identifier><identifier>EISSN: 1559-131X</identifier><identifier>EISSN: 1357-0560</identifier><identifier>DOI: 10.1385/MO:20:4:335</identifier><identifier>PMID: 14716029</identifier><identifier>CODEN: MONCEZ</identifier><language>eng</language><publisher>United States: Springer Nature B.V</publisher><subject>Dendritic Cells - immunology ; Dendritic Cells - metabolism ; Electroporation - methods ; Green Fluorescent Proteins ; Humans ; Immunotherapy - methods ; Leukemia ; Luminescent Proteins - metabolism ; Oncology ; RNA, Messenger - genetics ; RNA, Messenger - isolation &amp; purification ; Transfection - methods ; Tumor Cells, Cultured - metabolism</subject><ispartof>Medical oncology (Northwood, London, England), 2003-01, Vol.20 (4), p.335-348</ispartof><rights>Humana Press Inc. 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c312t-d9dcfa6d98c4662599ac123e527a061ea9389c90f938a1d2c94a10fa8985b7163</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,786,790,27957,27958</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14716029$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Takahashi, Masuhiro</creatorcontrib><creatorcontrib>Narita, Miwako</creatorcontrib><creatorcontrib>Ayres, Flavio</creatorcontrib><creatorcontrib>Satoh, Naoko</creatorcontrib><creatorcontrib>Abe, Takashi</creatorcontrib><creatorcontrib>Yanao, Toshio</creatorcontrib><creatorcontrib>Furukawa, Tatsuo</creatorcontrib><creatorcontrib>Toba, Ken</creatorcontrib><creatorcontrib>Hirohashi, Takeshi</creatorcontrib><creatorcontrib>Aizawa, Yoshifusa</creatorcontrib><title>Cytoplasmic expression of EGFP in dendritic cells transfected with in vitro transcribed mRNA or cellular total RNA extracted from EGFP expressing leukemia cells</title><title>Medical oncology (Northwood, London, England)</title><addtitle>Med Oncol</addtitle><description>The present study was designed for identifying the protein synthesis in cytoplasm of dendritic cells transfected with in vitro transcribed mRNA and cellular total RNA extracted from tumor cells. Dendritic cells were generated from cord blood-CD34+ cells by culture with GM-CSF, SCF, and TNF-alpha, or from peripheral blood adherent cells or CD14+ cells by culture with GM-CSF and IL-4. Dendritic cells were transfected with in vitro transcribed EGFP mRNA or cellular total RNA, which was isolated from EGFP expressing K562, by electroporation using a square-wave pulse. Optimal in vitro transcribed EGFP mRNA transfection efficiency (&gt;90%) was observed in a single electroporation of 1.75 kV/cm (electric field strength) with a pulse width of 250 micros. Although the intensity of EGFP expression in dendritic cells transfected with cellular total RNA was less compared with that in dendritic cells transfected with in vitro transcribed EGFP mRNA, a definite cytoplasmic synthesis of EGFP was demonstrated in dendritic cells transfected with cellular total RNA. The visual identification of cytoplasmic expression of cellular total RNA in dendritic cells revealed that electroporation of tumor cell-derived RNA could be a useful tool to load dendritic cells with tumor antigens for establishing an efficient dendritic cell-based tumor immunotherapy.</description><subject>Dendritic Cells - immunology</subject><subject>Dendritic Cells - metabolism</subject><subject>Electroporation - methods</subject><subject>Green Fluorescent Proteins</subject><subject>Humans</subject><subject>Immunotherapy - methods</subject><subject>Leukemia</subject><subject>Luminescent Proteins - metabolism</subject><subject>Oncology</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - isolation &amp; purification</subject><subject>Transfection - methods</subject><subject>Tumor Cells, Cultured - metabolism</subject><issn>1357-0560</issn><issn>1559-131X</issn><issn>1357-0560</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><recordid>eNpdkUlPwzAQhS0EYj9xRxYHLijgJU7i3qqqLBKlCIHEzXIdB1ySuNgOy7_hp-KSIiROY_l9M280D4ADjE4xLdjZZDogaJAOKGVrYBszxhNM8eN6fFOWJ4hlaAvseD9HiGBG-CbYwmmOM0T4NvgafQa7qKVvjIL6Y-G098a20FZwfHF-C00LS92WzoSoK13XHgYnW19pFXQJ3014XjJvJjjbK8qZWVSau5shtO6np6ulg8EGWcPlr_6I4E975WzT-_xat0-w1t2Lbozs7fbARiVrr_dXdRc8nI_vR5fJ9fTiajS8ThTFJCQlL1Uls5IXKs0ywjiXChOqGcklyrCWnBZccVTFKnFJFE8lRpUseMFm8Rh0Fxz3cxfOvnbaB9EYv9xAttp2XuSYIcJSGsGjf-Dcdq6Nu4kiZyzHhPAInfSQctZ7pyuxcKaR7lNgJJapiclUECRSEVOL9OFqZDdrdPnHrmKi35ZMk_E</recordid><startdate>20030101</startdate><enddate>20030101</enddate><creator>Takahashi, Masuhiro</creator><creator>Narita, Miwako</creator><creator>Ayres, Flavio</creator><creator>Satoh, Naoko</creator><creator>Abe, Takashi</creator><creator>Yanao, Toshio</creator><creator>Furukawa, Tatsuo</creator><creator>Toba, Ken</creator><creator>Hirohashi, Takeshi</creator><creator>Aizawa, Yoshifusa</creator><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope></search><sort><creationdate>20030101</creationdate><title>Cytoplasmic expression of EGFP in dendritic cells transfected with in vitro transcribed mRNA or cellular total RNA extracted from EGFP expressing leukemia cells</title><author>Takahashi, Masuhiro ; Narita, Miwako ; Ayres, Flavio ; Satoh, Naoko ; Abe, Takashi ; Yanao, Toshio ; Furukawa, Tatsuo ; Toba, Ken ; Hirohashi, Takeshi ; Aizawa, Yoshifusa</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c312t-d9dcfa6d98c4662599ac123e527a061ea9389c90f938a1d2c94a10fa8985b7163</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Dendritic Cells - immunology</topic><topic>Dendritic Cells - metabolism</topic><topic>Electroporation - methods</topic><topic>Green Fluorescent Proteins</topic><topic>Humans</topic><topic>Immunotherapy - methods</topic><topic>Leukemia</topic><topic>Luminescent Proteins - metabolism</topic><topic>Oncology</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - isolation &amp; purification</topic><topic>Transfection - methods</topic><topic>Tumor Cells, Cultured - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Takahashi, Masuhiro</creatorcontrib><creatorcontrib>Narita, Miwako</creatorcontrib><creatorcontrib>Ayres, Flavio</creatorcontrib><creatorcontrib>Satoh, Naoko</creatorcontrib><creatorcontrib>Abe, Takashi</creatorcontrib><creatorcontrib>Yanao, Toshio</creatorcontrib><creatorcontrib>Furukawa, Tatsuo</creatorcontrib><creatorcontrib>Toba, Ken</creatorcontrib><creatorcontrib>Hirohashi, Takeshi</creatorcontrib><creatorcontrib>Aizawa, Yoshifusa</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>ProQuest Health &amp; Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>Health &amp; Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>MEDLINE - Academic</collection><jtitle>Medical oncology (Northwood, London, England)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Takahashi, Masuhiro</au><au>Narita, Miwako</au><au>Ayres, Flavio</au><au>Satoh, Naoko</au><au>Abe, Takashi</au><au>Yanao, Toshio</au><au>Furukawa, Tatsuo</au><au>Toba, Ken</au><au>Hirohashi, Takeshi</au><au>Aizawa, Yoshifusa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cytoplasmic expression of EGFP in dendritic cells transfected with in vitro transcribed mRNA or cellular total RNA extracted from EGFP expressing leukemia cells</atitle><jtitle>Medical oncology (Northwood, London, England)</jtitle><addtitle>Med Oncol</addtitle><date>2003-01-01</date><risdate>2003</risdate><volume>20</volume><issue>4</issue><spage>335</spage><epage>348</epage><pages>335-348</pages><issn>1357-0560</issn><eissn>1559-131X</eissn><eissn>1357-0560</eissn><coden>MONCEZ</coden><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>The present study was designed for identifying the protein synthesis in cytoplasm of dendritic cells transfected with in vitro transcribed mRNA and cellular total RNA extracted from tumor cells. Dendritic cells were generated from cord blood-CD34+ cells by culture with GM-CSF, SCF, and TNF-alpha, or from peripheral blood adherent cells or CD14+ cells by culture with GM-CSF and IL-4. Dendritic cells were transfected with in vitro transcribed EGFP mRNA or cellular total RNA, which was isolated from EGFP expressing K562, by electroporation using a square-wave pulse. Optimal in vitro transcribed EGFP mRNA transfection efficiency (&gt;90%) was observed in a single electroporation of 1.75 kV/cm (electric field strength) with a pulse width of 250 micros. Although the intensity of EGFP expression in dendritic cells transfected with cellular total RNA was less compared with that in dendritic cells transfected with in vitro transcribed EGFP mRNA, a definite cytoplasmic synthesis of EGFP was demonstrated in dendritic cells transfected with cellular total RNA. The visual identification of cytoplasmic expression of cellular total RNA in dendritic cells revealed that electroporation of tumor cell-derived RNA could be a useful tool to load dendritic cells with tumor antigens for establishing an efficient dendritic cell-based tumor immunotherapy.</abstract><cop>United States</cop><pub>Springer Nature B.V</pub><pmid>14716029</pmid><doi>10.1385/MO:20:4:335</doi><tpages>14</tpages></addata></record>
fulltext fulltext
identifier ISSN: 1357-0560
ispartof Medical oncology (Northwood, London, England), 2003-01, Vol.20 (4), p.335-348
issn 1357-0560
1559-131X
1357-0560
language eng
recordid cdi_proquest_miscellaneous_71502543
source Springer Link
subjects Dendritic Cells - immunology
Dendritic Cells - metabolism
Electroporation - methods
Green Fluorescent Proteins
Humans
Immunotherapy - methods
Leukemia
Luminescent Proteins - metabolism
Oncology
RNA, Messenger - genetics
RNA, Messenger - isolation & purification
Transfection - methods
Tumor Cells, Cultured - metabolism
title Cytoplasmic expression of EGFP in dendritic cells transfected with in vitro transcribed mRNA or cellular total RNA extracted from EGFP expressing leukemia cells
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-09-21T23%3A30%3A57IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Cytoplasmic%20expression%20of%20EGFP%20in%20dendritic%20cells%20transfected%20with%20in%20vitro%20transcribed%20mRNA%20or%20cellular%20total%20RNA%20extracted%20from%20EGFP%20expressing%20leukemia%20cells&rft.jtitle=Medical%20oncology%20(Northwood,%20London,%20England)&rft.au=Takahashi,%20Masuhiro&rft.date=2003-01-01&rft.volume=20&rft.issue=4&rft.spage=335&rft.epage=348&rft.pages=335-348&rft.issn=1357-0560&rft.eissn=1559-131X&rft.coden=MONCEZ&rft_id=info:doi/10.1385/MO:20:4:335&rft_dat=%3Cproquest_cross%3E2394108251%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c312t-d9dcfa6d98c4662599ac123e527a061ea9389c90f938a1d2c94a10fa8985b7163%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=875571229&rft_id=info:pmid/14716029&rfr_iscdi=true