Long term expansion of undifferentiated human iPS and ES cells in suspension culture using a defined medium
Therapeutic application of stem cell derivatives requires large quantities of cells produced in defined media that cannot be produced via conventional adherent culture. We have applied human induced pluripotent stem (hiPS) cells expressing eGFP under control of the OCT4 promoter to establish the exp...
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| Veröffentlicht in: | Stem cell research 2010-07, Vol.5 (1), p.51-64 |
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| container_title | Stem cell research |
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| creator | Olmer, Ruth Haase, Alexandra Merkert, Sylvia Cui, Wei Paleček, Jiří Ran, Chen Kirschning, Andreas Scheper, Thomas Glage, Silke Miller, Konstantin Curnow, Eliza C. Hayes, Eric S. Martin, Ulrich |
| description | Therapeutic application of stem cell derivatives requires large quantities of cells produced in defined media that cannot be produced via conventional adherent culture. We have applied human induced pluripotent stem (hiPS) cells expressing eGFP under control of the OCT4 promoter to establish the expansion of undifferentiated human embryonic stem (hES) and hiPS cells in suspension culture. A defined culture medium has been identified that results in up to six-fold increase in cell numbers within four days. Our culture system is based on initial single cell dissociation which is critical for standardized process inoculation. HES / hiPS cells were expanded for up to 17 passages. The cells maintained a stable karyotype, their expression of pluripotency markers and their potential to differentiate into derivatives of all three germ layers. The ability to expand HES / hiPS cells in a scalable suspension culture represents a critical step towards standardized production in stirred bioreactors. |
| doi_str_mv | 10.1016/j.scr.2010.03.005 |
| format | Article |
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We have applied human induced pluripotent stem (hiPS) cells expressing eGFP under control of the OCT4 promoter to establish the expansion of undifferentiated human embryonic stem (hES) and hiPS cells in suspension culture. A defined culture medium has been identified that results in up to six-fold increase in cell numbers within four days. Our culture system is based on initial single cell dissociation which is critical for standardized process inoculation. HES / hiPS cells were expanded for up to 17 passages. The cells maintained a stable karyotype, their expression of pluripotency markers and their potential to differentiate into derivatives of all three germ layers. The ability to expand HES / hiPS cells in a scalable suspension culture represents a critical step towards standardized production in stirred bioreactors.</description><identifier>ISSN: 1873-5061</identifier><identifier>EISSN: 1876-7753</identifier><identifier>DOI: 10.1016/j.scr.2010.03.005</identifier><identifier>PMID: 20478754</identifier><language>eng</language><publisher>England: Elsevier B.V</publisher><subject>Cell Culture Techniques - methods ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Culture Media ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - metabolism ; Humans ; Induced Pluripotent Stem Cells - cytology ; Induced Pluripotent Stem Cells - metabolism ; Octamer Transcription Factor-3 - genetics ; Promoter Regions, Genetic ; Time Factors</subject><ispartof>Stem cell research, 2010-07, Vol.5 (1), p.51-64</ispartof><rights>2010 Elsevier B.V.</rights><rights>Copyright 2010 Elsevier B.V. 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We have applied human induced pluripotent stem (hiPS) cells expressing eGFP under control of the OCT4 promoter to establish the expansion of undifferentiated human embryonic stem (hES) and hiPS cells in suspension culture. A defined culture medium has been identified that results in up to six-fold increase in cell numbers within four days. Our culture system is based on initial single cell dissociation which is critical for standardized process inoculation. HES / hiPS cells were expanded for up to 17 passages. The cells maintained a stable karyotype, their expression of pluripotency markers and their potential to differentiate into derivatives of all three germ layers. The ability to expand HES / hiPS cells in a scalable suspension culture represents a critical step towards standardized production in stirred bioreactors.</description><subject>Cell Culture Techniques - methods</subject><subject>Cell Differentiation</subject><subject>Cell Line</subject><subject>Cell Proliferation</subject><subject>Culture Media</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - metabolism</subject><subject>Humans</subject><subject>Induced Pluripotent Stem Cells - cytology</subject><subject>Induced Pluripotent Stem Cells - metabolism</subject><subject>Octamer Transcription Factor-3 - genetics</subject><subject>Promoter Regions, Genetic</subject><subject>Time Factors</subject><issn>1873-5061</issn><issn>1876-7753</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1PxCAURYnR-P0D3Bh2rjpCgULjypjxI5lEE3VNEF6VcUpHKEb_vYyjLmUDJPeevHcQOqJkQgltTueTZOOkJuVP2IQQsYF2qZJNJaVgm99vVgnS0B20l9K8BNpa1dtopyZcKin4LnqdDeEZjxB7DB9LE5IfAh46nIPzXQcRwujNCA6_5N4E7O_usQkOT--xhcUiYR9wymkJ66LNizFHwDn5QjXYQedDKffgfO4P0FZnFgkOf-599Hg5fbi4rma3VzcX57PKcqrGyrWdakXTCJBWcU4I74RshJWqEdw48WR53XLneC1rA2ANOMcax1pVt5YbxvbRyZq7jMNbhjTq3qfVuCbAkJOWrBxFOC1Juk7aOKQUodPL6HsTPzUleqVYz3VRrFeKNWG6GCyd4x96fip7_TV-nZbA2ToAZcd3D7EgPARbHESwo3aD_wf_BZ6njPw</recordid><startdate>20100701</startdate><enddate>20100701</enddate><creator>Olmer, Ruth</creator><creator>Haase, Alexandra</creator><creator>Merkert, Sylvia</creator><creator>Cui, Wei</creator><creator>Paleček, Jiří</creator><creator>Ran, Chen</creator><creator>Kirschning, Andreas</creator><creator>Scheper, Thomas</creator><creator>Glage, Silke</creator><creator>Miller, Konstantin</creator><creator>Curnow, Eliza C.</creator><creator>Hayes, Eric S.</creator><creator>Martin, Ulrich</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</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>7X8</scope></search><sort><creationdate>20100701</creationdate><title>Long term expansion of undifferentiated human iPS and ES cells in suspension culture using a defined medium</title><author>Olmer, Ruth ; Haase, Alexandra ; Merkert, Sylvia ; Cui, Wei ; Paleček, Jiří ; Ran, Chen ; Kirschning, Andreas ; Scheper, Thomas ; Glage, Silke ; Miller, Konstantin ; Curnow, Eliza C. ; Hayes, Eric S. ; Martin, Ulrich</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c418t-d9f895665e7c844004f5765c78654ad5bc4294dd4272aeecaedd36d39829c4a33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Cell Culture Techniques - methods</topic><topic>Cell Differentiation</topic><topic>Cell Line</topic><topic>Cell Proliferation</topic><topic>Culture Media</topic><topic>Embryonic Stem Cells - cytology</topic><topic>Embryonic Stem Cells - metabolism</topic><topic>Humans</topic><topic>Induced Pluripotent Stem Cells - cytology</topic><topic>Induced Pluripotent Stem Cells - metabolism</topic><topic>Octamer Transcription Factor-3 - genetics</topic><topic>Promoter Regions, Genetic</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Olmer, Ruth</creatorcontrib><creatorcontrib>Haase, Alexandra</creatorcontrib><creatorcontrib>Merkert, Sylvia</creatorcontrib><creatorcontrib>Cui, Wei</creatorcontrib><creatorcontrib>Paleček, Jiří</creatorcontrib><creatorcontrib>Ran, Chen</creatorcontrib><creatorcontrib>Kirschning, Andreas</creatorcontrib><creatorcontrib>Scheper, Thomas</creatorcontrib><creatorcontrib>Glage, Silke</creatorcontrib><creatorcontrib>Miller, Konstantin</creatorcontrib><creatorcontrib>Curnow, Eliza C.</creatorcontrib><creatorcontrib>Hayes, Eric S.</creatorcontrib><creatorcontrib>Martin, Ulrich</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Stem cell research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Olmer, Ruth</au><au>Haase, Alexandra</au><au>Merkert, Sylvia</au><au>Cui, Wei</au><au>Paleček, Jiří</au><au>Ran, Chen</au><au>Kirschning, Andreas</au><au>Scheper, Thomas</au><au>Glage, Silke</au><au>Miller, Konstantin</au><au>Curnow, Eliza C.</au><au>Hayes, Eric S.</au><au>Martin, Ulrich</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Long term expansion of undifferentiated human iPS and ES cells in suspension culture using a defined medium</atitle><jtitle>Stem cell research</jtitle><addtitle>Stem Cell Res</addtitle><date>2010-07-01</date><risdate>2010</risdate><volume>5</volume><issue>1</issue><spage>51</spage><epage>64</epage><pages>51-64</pages><issn>1873-5061</issn><eissn>1876-7753</eissn><abstract>Therapeutic application of stem cell derivatives requires large quantities of cells produced in defined media that cannot be produced via conventional adherent culture. 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| ispartof | Stem cell research, 2010-07, Vol.5 (1), p.51-64 |
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| language | eng |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present); EZB-FREE-00999 freely available EZB journals |
| subjects | Cell Culture Techniques - methods Cell Differentiation Cell Line Cell Proliferation Culture Media Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Humans Induced Pluripotent Stem Cells - cytology Induced Pluripotent Stem Cells - metabolism Octamer Transcription Factor-3 - genetics Promoter Regions, Genetic Time Factors |
| title | Long term expansion of undifferentiated human iPS and ES cells in suspension culture using a defined medium |
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