A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use
Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required t...
Gespeichert in:
Veröffentlicht in: | Journal of immunological methods 2000-11, Vol.245 (1), p.15-29 |
---|---|
Hauptverfasser: | , , , , , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 29 |
---|---|
container_issue | 1 |
container_start_page | 15 |
container_title | Journal of immunological methods |
container_volume | 245 |
creator | Feuerstein, Bernadette Berger, Thomas G Maczek, Christian Röder, Claudia Schreiner, Doris Hirsch, Ute Haendle, Ina Leisgang, Waltraud Glaser, Anke Kuss, Oliver Diepgen, Thomas L Schuler, Gerold Schuler-Thurner, Beatrice |
description | Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aimed at developing an effective cryopreservation method, and by testing several variables found that it was crucial to combine the most advantageous maturation stimulus with an improved freezing procedure. We generated monocyte-derived DC from leukapheresis products by using GM–CSF and IL-4 and showed that amongst several known maturation stimuli the cocktail consisting of TNF-alpha+IL-1 beta+IL-6+PGE
2 achieved the highest survival of mature DC. We then systematically explored cryopreservation conditions, and found that freezing matured DC at 1°C/min in pure autologous serum+10% DMSO+5% glucose at a cell density of 10×10
6 DC/ml gave the best results. Using this approach 85–100% of the frozen DC could be recovered in a viable state after thawing (
Table 1). The morphology, phenotype, survival as well as functional properties (allogeneic mixed leukocyte reaction, induction of influenza matrix or melan A peptide-specific cytotoxic T cells) of these thawed DC were equivalent to freshly prepared ones. The addition of CD40L or TRANCE/RANKL further improved DC survival. Importantly, we demonstrate that DC can effectively be loaded with antigens (such as Tetanus Toxoid, influenza matrix and melan A peptides) before cryopreservation so that it is now possible to generate antigen-preloaded, frozen DC aliquots that after thawing can be used right away. This is an important advance as both the generation of a standardized DC vaccine under GMP conditions and the carrying out of clinical trials are greatly facilitated. |
doi_str_mv | 10.1016/S0022-1759(00)00269-6 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_72355292</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0022175900002696</els_id><sourcerecordid>72355292</sourcerecordid><originalsourceid>FETCH-LOGICAL-c488t-43e19125276bd92b0c04303c3ffd040f047fda5d5ba6bb7c416ec82f3f74f1c03</originalsourceid><addsrcrecordid>eNqFkUFv1DAQhS0EotvCTwD5AgKJwNhO4uSEqqq0lSr1AJwtxx5ToyTe2k6l_fc4uyt67MkazTfjN-8R8o7BVwas_fYTgPOKyab_BPC5FG1ftS_IhnWSV7KH5iXZ_EdOyGlKfwGAQQuvyQljUHPewYbszumE-T5Y6kKk-R7pNga7mOzDTIOjJu7CNmLC-IiW6tE_LCGntaPn7P_gXJXuGLRF-4VOOi8RqcXZRp-9oQbHMdGI2u72-83oZ2_0SJeEb8grp8eEb4_vGfn94_LXxXV1e3d1c3F-W5m663JVC2Q94w2X7WB7PoCBWoAwwjkLNTiopbO6sc2g22GQpmYtmo474WTtmAFxRj4e9pbDHhZMWU0-rcL0jGFJSnLRNLznz4JMSg6ibwrYHEATQ0oRndpGP-m4UwzUGo7ah6NW5xWA2oej2jL3_vjBMkxon6aOaRTgwxHQqbjkop6NT09cuY0Jtgr4fuCw-PboMapkPM4GrY9osrLBPyPlH4acrM0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17720395</pqid></control><display><type>article</type><title>A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Feuerstein, Bernadette ; Berger, Thomas G ; Maczek, Christian ; Röder, Claudia ; Schreiner, Doris ; Hirsch, Ute ; Haendle, Ina ; Leisgang, Waltraud ; Glaser, Anke ; Kuss, Oliver ; Diepgen, Thomas L ; Schuler, Gerold ; Schuler-Thurner, Beatrice</creator><creatorcontrib>Feuerstein, Bernadette ; Berger, Thomas G ; Maczek, Christian ; Röder, Claudia ; Schreiner, Doris ; Hirsch, Ute ; Haendle, Ina ; Leisgang, Waltraud ; Glaser, Anke ; Kuss, Oliver ; Diepgen, Thomas L ; Schuler, Gerold ; Schuler-Thurner, Beatrice</creatorcontrib><description>Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aimed at developing an effective cryopreservation method, and by testing several variables found that it was crucial to combine the most advantageous maturation stimulus with an improved freezing procedure. We generated monocyte-derived DC from leukapheresis products by using GM–CSF and IL-4 and showed that amongst several known maturation stimuli the cocktail consisting of TNF-alpha+IL-1 beta+IL-6+PGE
2 achieved the highest survival of mature DC. We then systematically explored cryopreservation conditions, and found that freezing matured DC at 1°C/min in pure autologous serum+10% DMSO+5% glucose at a cell density of 10×10
6 DC/ml gave the best results. Using this approach 85–100% of the frozen DC could be recovered in a viable state after thawing (
Table 1). The morphology, phenotype, survival as well as functional properties (allogeneic mixed leukocyte reaction, induction of influenza matrix or melan A peptide-specific cytotoxic T cells) of these thawed DC were equivalent to freshly prepared ones. The addition of CD40L or TRANCE/RANKL further improved DC survival. Importantly, we demonstrate that DC can effectively be loaded with antigens (such as Tetanus Toxoid, influenza matrix and melan A peptides) before cryopreservation so that it is now possible to generate antigen-preloaded, frozen DC aliquots that after thawing can be used right away. This is an important advance as both the generation of a standardized DC vaccine under GMP conditions and the carrying out of clinical trials are greatly facilitated.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/S0022-1759(00)00269-6</identifier><identifier>PMID: 11042280</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject><![CDATA[Antigens - administration & dosage ; Biological and medical sciences ; Carrier Proteins - administration & dosage ; CD40 Ligand - administration & dosage ; Cell Differentiation ; Cell Survival ; Cryopreservation ; Cryopreservation - methods ; Dendritic cells ; Dendritic Cells - cytology ; Dendritic Cells - immunology ; Diverse techniques ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Humans ; Immunobiology ; Immunotherapy ; In Vitro Techniques ; Lymphocyte Activation ; Lymphocyte Culture Test, Mixed ; Lymphoid cells: ontogeny, maturation, markers, receptors, circulation and recirculation ; Membrane Glycoproteins - administration & dosage ; Microbiology ; Molecular and cellular biology ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; T-Lymphocytes, Cytotoxic - immunology ; Tetanus Toxoid - administration & dosage ; Vaccines - administration & dosage]]></subject><ispartof>Journal of immunological methods, 2000-11, Vol.245 (1), p.15-29</ispartof><rights>2000 Elsevier Science B.V.</rights><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c488t-43e19125276bd92b0c04303c3ffd040f047fda5d5ba6bb7c416ec82f3f74f1c03</citedby><cites>FETCH-LOGICAL-c488t-43e19125276bd92b0c04303c3ffd040f047fda5d5ba6bb7c416ec82f3f74f1c03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0022-1759(00)00269-6$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14161315$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11042280$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Feuerstein, Bernadette</creatorcontrib><creatorcontrib>Berger, Thomas G</creatorcontrib><creatorcontrib>Maczek, Christian</creatorcontrib><creatorcontrib>Röder, Claudia</creatorcontrib><creatorcontrib>Schreiner, Doris</creatorcontrib><creatorcontrib>Hirsch, Ute</creatorcontrib><creatorcontrib>Haendle, Ina</creatorcontrib><creatorcontrib>Leisgang, Waltraud</creatorcontrib><creatorcontrib>Glaser, Anke</creatorcontrib><creatorcontrib>Kuss, Oliver</creatorcontrib><creatorcontrib>Diepgen, Thomas L</creatorcontrib><creatorcontrib>Schuler, Gerold</creatorcontrib><creatorcontrib>Schuler-Thurner, Beatrice</creatorcontrib><title>A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aimed at developing an effective cryopreservation method, and by testing several variables found that it was crucial to combine the most advantageous maturation stimulus with an improved freezing procedure. We generated monocyte-derived DC from leukapheresis products by using GM–CSF and IL-4 and showed that amongst several known maturation stimuli the cocktail consisting of TNF-alpha+IL-1 beta+IL-6+PGE
2 achieved the highest survival of mature DC. We then systematically explored cryopreservation conditions, and found that freezing matured DC at 1°C/min in pure autologous serum+10% DMSO+5% glucose at a cell density of 10×10
6 DC/ml gave the best results. Using this approach 85–100% of the frozen DC could be recovered in a viable state after thawing (
Table 1). The morphology, phenotype, survival as well as functional properties (allogeneic mixed leukocyte reaction, induction of influenza matrix or melan A peptide-specific cytotoxic T cells) of these thawed DC were equivalent to freshly prepared ones. The addition of CD40L or TRANCE/RANKL further improved DC survival. Importantly, we demonstrate that DC can effectively be loaded with antigens (such as Tetanus Toxoid, influenza matrix and melan A peptides) before cryopreservation so that it is now possible to generate antigen-preloaded, frozen DC aliquots that after thawing can be used right away. This is an important advance as both the generation of a standardized DC vaccine under GMP conditions and the carrying out of clinical trials are greatly facilitated.</description><subject>Antigens - administration & dosage</subject><subject>Biological and medical sciences</subject><subject>Carrier Proteins - administration & dosage</subject><subject>CD40 Ligand - administration & dosage</subject><subject>Cell Differentiation</subject><subject>Cell Survival</subject><subject>Cryopreservation</subject><subject>Cryopreservation - methods</subject><subject>Dendritic cells</subject><subject>Dendritic Cells - cytology</subject><subject>Dendritic Cells - immunology</subject><subject>Diverse techniques</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Humans</subject><subject>Immunobiology</subject><subject>Immunotherapy</subject><subject>In Vitro Techniques</subject><subject>Lymphocyte Activation</subject><subject>Lymphocyte Culture Test, Mixed</subject><subject>Lymphoid cells: ontogeny, maturation, markers, receptors, circulation and recirculation</subject><subject>Membrane Glycoproteins - administration & dosage</subject><subject>Microbiology</subject><subject>Molecular and cellular biology</subject><subject>RANK Ligand</subject><subject>Receptor Activator of Nuclear Factor-kappa B</subject><subject>T-Lymphocytes, Cytotoxic - immunology</subject><subject>Tetanus Toxoid - administration & dosage</subject><subject>Vaccines - administration & dosage</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUFv1DAQhS0EotvCTwD5AgKJwNhO4uSEqqq0lSr1AJwtxx5ToyTe2k6l_fc4uyt67MkazTfjN-8R8o7BVwas_fYTgPOKyab_BPC5FG1ftS_IhnWSV7KH5iXZ_EdOyGlKfwGAQQuvyQljUHPewYbszumE-T5Y6kKk-R7pNga7mOzDTIOjJu7CNmLC-IiW6tE_LCGntaPn7P_gXJXuGLRF-4VOOi8RqcXZRp-9oQbHMdGI2u72-83oZ2_0SJeEb8grp8eEb4_vGfn94_LXxXV1e3d1c3F-W5m663JVC2Q94w2X7WB7PoCBWoAwwjkLNTiopbO6sc2g22GQpmYtmo474WTtmAFxRj4e9pbDHhZMWU0-rcL0jGFJSnLRNLznz4JMSg6ibwrYHEATQ0oRndpGP-m4UwzUGo7ah6NW5xWA2oej2jL3_vjBMkxon6aOaRTgwxHQqbjkop6NT09cuY0Jtgr4fuCw-PboMapkPM4GrY9osrLBPyPlH4acrM0</recordid><startdate>20001101</startdate><enddate>20001101</enddate><creator>Feuerstein, Bernadette</creator><creator>Berger, Thomas G</creator><creator>Maczek, Christian</creator><creator>Röder, Claudia</creator><creator>Schreiner, Doris</creator><creator>Hirsch, Ute</creator><creator>Haendle, Ina</creator><creator>Leisgang, Waltraud</creator><creator>Glaser, Anke</creator><creator>Kuss, Oliver</creator><creator>Diepgen, Thomas L</creator><creator>Schuler, Gerold</creator><creator>Schuler-Thurner, Beatrice</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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>7QO</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20001101</creationdate><title>A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use</title><author>Feuerstein, Bernadette ; Berger, Thomas G ; Maczek, Christian ; Röder, Claudia ; Schreiner, Doris ; Hirsch, Ute ; Haendle, Ina ; Leisgang, Waltraud ; Glaser, Anke ; Kuss, Oliver ; Diepgen, Thomas L ; Schuler, Gerold ; Schuler-Thurner, Beatrice</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c488t-43e19125276bd92b0c04303c3ffd040f047fda5d5ba6bb7c416ec82f3f74f1c03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Antigens - administration & dosage</topic><topic>Biological and medical sciences</topic><topic>Carrier Proteins - administration & dosage</topic><topic>CD40 Ligand - administration & dosage</topic><topic>Cell Differentiation</topic><topic>Cell Survival</topic><topic>Cryopreservation</topic><topic>Cryopreservation - methods</topic><topic>Dendritic cells</topic><topic>Dendritic Cells - cytology</topic><topic>Dendritic Cells - immunology</topic><topic>Diverse techniques</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Humans</topic><topic>Immunobiology</topic><topic>Immunotherapy</topic><topic>In Vitro Techniques</topic><topic>Lymphocyte Activation</topic><topic>Lymphocyte Culture Test, Mixed</topic><topic>Lymphoid cells: ontogeny, maturation, markers, receptors, circulation and recirculation</topic><topic>Membrane Glycoproteins - administration & dosage</topic><topic>Microbiology</topic><topic>Molecular and cellular biology</topic><topic>RANK Ligand</topic><topic>Receptor Activator of Nuclear Factor-kappa B</topic><topic>T-Lymphocytes, Cytotoxic - immunology</topic><topic>Tetanus Toxoid - administration & dosage</topic><topic>Vaccines - administration & dosage</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Feuerstein, Bernadette</creatorcontrib><creatorcontrib>Berger, Thomas G</creatorcontrib><creatorcontrib>Maczek, Christian</creatorcontrib><creatorcontrib>Röder, Claudia</creatorcontrib><creatorcontrib>Schreiner, Doris</creatorcontrib><creatorcontrib>Hirsch, Ute</creatorcontrib><creatorcontrib>Haendle, Ina</creatorcontrib><creatorcontrib>Leisgang, Waltraud</creatorcontrib><creatorcontrib>Glaser, Anke</creatorcontrib><creatorcontrib>Kuss, Oliver</creatorcontrib><creatorcontrib>Diepgen, Thomas L</creatorcontrib><creatorcontrib>Schuler, Gerold</creatorcontrib><creatorcontrib>Schuler-Thurner, Beatrice</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Feuerstein, Bernadette</au><au>Berger, Thomas G</au><au>Maczek, Christian</au><au>Röder, Claudia</au><au>Schreiner, Doris</au><au>Hirsch, Ute</au><au>Haendle, Ina</au><au>Leisgang, Waltraud</au><au>Glaser, Anke</au><au>Kuss, Oliver</au><au>Diepgen, Thomas L</au><au>Schuler, Gerold</au><au>Schuler-Thurner, Beatrice</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>2000-11-01</date><risdate>2000</risdate><volume>245</volume><issue>1</issue><spage>15</spage><epage>29</epage><pages>15-29</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aimed at developing an effective cryopreservation method, and by testing several variables found that it was crucial to combine the most advantageous maturation stimulus with an improved freezing procedure. We generated monocyte-derived DC from leukapheresis products by using GM–CSF and IL-4 and showed that amongst several known maturation stimuli the cocktail consisting of TNF-alpha+IL-1 beta+IL-6+PGE
2 achieved the highest survival of mature DC. We then systematically explored cryopreservation conditions, and found that freezing matured DC at 1°C/min in pure autologous serum+10% DMSO+5% glucose at a cell density of 10×10
6 DC/ml gave the best results. Using this approach 85–100% of the frozen DC could be recovered in a viable state after thawing (
Table 1). The morphology, phenotype, survival as well as functional properties (allogeneic mixed leukocyte reaction, induction of influenza matrix or melan A peptide-specific cytotoxic T cells) of these thawed DC were equivalent to freshly prepared ones. The addition of CD40L or TRANCE/RANKL further improved DC survival. Importantly, we demonstrate that DC can effectively be loaded with antigens (such as Tetanus Toxoid, influenza matrix and melan A peptides) before cryopreservation so that it is now possible to generate antigen-preloaded, frozen DC aliquots that after thawing can be used right away. This is an important advance as both the generation of a standardized DC vaccine under GMP conditions and the carrying out of clinical trials are greatly facilitated.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>11042280</pmid><doi>10.1016/S0022-1759(00)00269-6</doi><tpages>15</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0022-1759 |
ispartof | Journal of immunological methods, 2000-11, Vol.245 (1), p.15-29 |
issn | 0022-1759 1872-7905 |
language | eng |
recordid | cdi_proquest_miscellaneous_72355292 |
source | MEDLINE; Access via ScienceDirect (Elsevier) |
subjects | Antigens - administration & dosage Biological and medical sciences Carrier Proteins - administration & dosage CD40 Ligand - administration & dosage Cell Differentiation Cell Survival Cryopreservation Cryopreservation - methods Dendritic cells Dendritic Cells - cytology Dendritic Cells - immunology Diverse techniques Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Immunobiology Immunotherapy In Vitro Techniques Lymphocyte Activation Lymphocyte Culture Test, Mixed Lymphoid cells: ontogeny, maturation, markers, receptors, circulation and recirculation Membrane Glycoproteins - administration & dosage Microbiology Molecular and cellular biology RANK Ligand Receptor Activator of Nuclear Factor-kappa B T-Lymphocytes, Cytotoxic - immunology Tetanus Toxoid - administration & dosage Vaccines - administration & dosage |
title | A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T04%3A44%3A37IST&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=A%20method%20for%20the%20production%20of%20cryopreserved%20aliquots%20of%20antigen-preloaded,%20mature%20dendritic%20cells%20ready%20for%20clinical%20use&rft.jtitle=Journal%20of%20immunological%20methods&rft.au=Feuerstein,%20Bernadette&rft.date=2000-11-01&rft.volume=245&rft.issue=1&rft.spage=15&rft.epage=29&rft.pages=15-29&rft.issn=0022-1759&rft.eissn=1872-7905&rft.coden=JIMMBG&rft_id=info:doi/10.1016/S0022-1759(00)00269-6&rft_dat=%3Cproquest_cross%3E72355292%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17720395&rft_id=info:pmid/11042280&rft_els_id=S0022175900002696&rfr_iscdi=true |