Rapid serum‐free/suspension adaptation: Medium development using a definitive screening design for Chinese hamster ovary cells
The biopharmaceutical industry prefers to culture the mammalian cells in suspension with a serum‐free media (SFM) due to improved productivity and process consistency. However, mammalian cells preferentially grow as adherent cells in a complete medium (CM) containing serum. Therefore, cells require...
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| Veröffentlicht in: | Biotechnology progress 2021-07, Vol.37 (4), p.e3154-n/a |
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| creator | Wu, Suyang Rish, Adam J. Skomo, Alec Zhao, Yuxiang Drennen, James K. Anderson, Carl A. |
| description | The biopharmaceutical industry prefers to culture the mammalian cells in suspension with a serum‐free media (SFM) due to improved productivity and process consistency. However, mammalian cells preferentially grow as adherent cells in a complete medium (CM) containing serum. Therefore, cells require adaptation from adherence in CM to suspension culture in SFM. This work proposes an adaptation method that includes media supplementation during the adaption of Chinese hamster ovary cells. As a result, the adaptation was accelerated compared to the traditional repetitive subculturing. Ca2+/Mg2+ supplementation significantly reduced the doubling time compared to the adaptation without supplementation during the adaptation of adherent cells from 100% CM to 75% CM (p |
| doi_str_mv | 10.1002/btpr.3154 |
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However, mammalian cells preferentially grow as adherent cells in a complete medium (CM) containing serum. Therefore, cells require adaptation from adherence in CM to suspension culture in SFM. This work proposes an adaptation method that includes media supplementation during the adaption of Chinese hamster ovary cells. As a result, the adaptation was accelerated compared to the traditional repetitive subculturing. Ca2+/Mg2+ supplementation significantly reduced the doubling time compared to the adaptation without supplementation during the adaptation of adherent cells from 100% CM to 75% CM (p < 0.05). Furthermore, a definitive screening design (DSD) was applied to select essential nutrients during the adaptation from 10% CM to 0% CM. The main effects of Ca2+ and Dulbecco's modified essential medium (DMEM) were found significant to both viable cell density and viability at harvest. Additionally, the interaction term between Ca2+ and DMEM was found significant, which highlights the ability of DSD to capture interaction terms. Eventually, the media supplementation method resulted in adaptation SFM in 27 days, compared to the previously reported 66 days. Additionally, the membrane surface integrin expression was found significantly decreased when adherent cells were adapted to suspension. Moreover, the Ca2+/Mg2+ supplementation correlated with faster integrin recovery after trypsinization. However, faster integrin recovery did not contribute to the accelerated cell growth when subculturing from 100% CM to 75% CM.</description><identifier>ISSN: 8756-7938</identifier><identifier>EISSN: 1520-6033</identifier><identifier>DOI: 10.1002/btpr.3154</identifier><identifier>PMID: 33864359</identifier><language>eng</language><publisher>Hoboken, USA: John Wiley & Sons, Inc</publisher><subject>Adaptation ; Adherent cells ; Biopharmaceuticals ; Calcium ; Calcium ions ; Cell culture ; Cell density ; CHO cells ; definitive screening design ; Essential nutrients ; integrin ; Magnesium ; Mammalian cells ; Mammals ; Nutrients ; Ovaries ; Recovery ; Screening ; serum‐free/suspension adaptation ; Supplements ; Suspension culture</subject><ispartof>Biotechnology progress, 2021-07, Vol.37 (4), p.e3154-n/a</ispartof><rights>2021 American Institute of Chemical Engineers</rights><rights>2021 American Institute of Chemical Engineers.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2504-6f12df02889fdcce12e568a5a4403e77350e4db441d72c726286319658f3fe8c3</citedby><cites>FETCH-LOGICAL-c2504-6f12df02889fdcce12e568a5a4403e77350e4db441d72c726286319658f3fe8c3</cites><orcidid>0000-0001-7069-6777</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbtpr.3154$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbtpr.3154$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33864359$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wu, Suyang</creatorcontrib><creatorcontrib>Rish, Adam J.</creatorcontrib><creatorcontrib>Skomo, Alec</creatorcontrib><creatorcontrib>Zhao, Yuxiang</creatorcontrib><creatorcontrib>Drennen, James K.</creatorcontrib><creatorcontrib>Anderson, Carl A.</creatorcontrib><title>Rapid serum‐free/suspension adaptation: Medium development using a definitive screening design for Chinese hamster ovary cells</title><title>Biotechnology progress</title><addtitle>Biotechnol Prog</addtitle><description>The biopharmaceutical industry prefers to culture the mammalian cells in suspension with a serum‐free media (SFM) due to improved productivity and process consistency. However, mammalian cells preferentially grow as adherent cells in a complete medium (CM) containing serum. Therefore, cells require adaptation from adherence in CM to suspension culture in SFM. This work proposes an adaptation method that includes media supplementation during the adaption of Chinese hamster ovary cells. As a result, the adaptation was accelerated compared to the traditional repetitive subculturing. Ca2+/Mg2+ supplementation significantly reduced the doubling time compared to the adaptation without supplementation during the adaptation of adherent cells from 100% CM to 75% CM (p < 0.05). Furthermore, a definitive screening design (DSD) was applied to select essential nutrients during the adaptation from 10% CM to 0% CM. The main effects of Ca2+ and Dulbecco's modified essential medium (DMEM) were found significant to both viable cell density and viability at harvest. Additionally, the interaction term between Ca2+ and DMEM was found significant, which highlights the ability of DSD to capture interaction terms. Eventually, the media supplementation method resulted in adaptation SFM in 27 days, compared to the previously reported 66 days. Additionally, the membrane surface integrin expression was found significantly decreased when adherent cells were adapted to suspension. Moreover, the Ca2+/Mg2+ supplementation correlated with faster integrin recovery after trypsinization. However, faster integrin recovery did not contribute to the accelerated cell growth when subculturing from 100% CM to 75% CM.</description><subject>Adaptation</subject><subject>Adherent cells</subject><subject>Biopharmaceuticals</subject><subject>Calcium</subject><subject>Calcium ions</subject><subject>Cell culture</subject><subject>Cell density</subject><subject>CHO cells</subject><subject>definitive screening design</subject><subject>Essential nutrients</subject><subject>integrin</subject><subject>Magnesium</subject><subject>Mammalian cells</subject><subject>Mammals</subject><subject>Nutrients</subject><subject>Ovaries</subject><subject>Recovery</subject><subject>Screening</subject><subject>serum‐free/suspension adaptation</subject><subject>Supplements</subject><subject>Suspension culture</subject><issn>8756-7938</issn><issn>1520-6033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNp1kMtKxDAUhoMoOl4WvoAEXLmok3sz7nTwBooiui6Z5kQjbVqTdsSdj-Az-iR2HHXn6hx-Pr5z-BHapeSQEsLGs66Nh5xKsYJGVDKSKcL5KhrpXKosn3C9gTZTeiaEaKLYOtrgXCvB5WSE3u9M6y1OEPv68_3DRYBx6lMLIfkmYGNN25luWI_wNVjf19jCHKqmrSF0uE8-PGIzZM4H3_k54FQOirCILST_GLBrIp4--QAJ8JOpUwcRN3MT33AJVZW20ZozVYKdn7mFHs5O76cX2dXN-eX0-CormSQiU44y6wjTeuJsWQJlIJU20ghBOOQ5lwSEnQlBbc7KnCmmFacTJbXjDnTJt9D-0tvG5qWH1BXPTR_DcLJgUhEmBRF0oA6WVBmblCK4oo2-Hp4tKCkWXReLrotF1wO792PsZzXYP_K33AEYL4FXX8Hb_6bi5P727lv5BQ_fi_Q</recordid><startdate>202107</startdate><enddate>202107</enddate><creator>Wu, Suyang</creator><creator>Rish, Adam J.</creator><creator>Skomo, Alec</creator><creator>Zhao, Yuxiang</creator><creator>Drennen, James K.</creator><creator>Anderson, Carl A.</creator><general>John Wiley & Sons, Inc</general><general>Wiley Subscription Services, Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><orcidid>https://orcid.org/0000-0001-7069-6777</orcidid></search><sort><creationdate>202107</creationdate><title>Rapid serum‐free/suspension adaptation: Medium development using a definitive screening design for Chinese hamster ovary cells</title><author>Wu, Suyang ; Rish, Adam J. ; Skomo, Alec ; Zhao, Yuxiang ; Drennen, James K. ; Anderson, Carl A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2504-6f12df02889fdcce12e568a5a4403e77350e4db441d72c726286319658f3fe8c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Adaptation</topic><topic>Adherent cells</topic><topic>Biopharmaceuticals</topic><topic>Calcium</topic><topic>Calcium ions</topic><topic>Cell culture</topic><topic>Cell density</topic><topic>CHO cells</topic><topic>definitive screening design</topic><topic>Essential nutrients</topic><topic>integrin</topic><topic>Magnesium</topic><topic>Mammalian cells</topic><topic>Mammals</topic><topic>Nutrients</topic><topic>Ovaries</topic><topic>Recovery</topic><topic>Screening</topic><topic>serum‐free/suspension adaptation</topic><topic>Supplements</topic><topic>Suspension culture</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wu, Suyang</creatorcontrib><creatorcontrib>Rish, Adam J.</creatorcontrib><creatorcontrib>Skomo, Alec</creatorcontrib><creatorcontrib>Zhao, Yuxiang</creatorcontrib><creatorcontrib>Drennen, James K.</creatorcontrib><creatorcontrib>Anderson, Carl A.</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Biotechnology progress</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wu, Suyang</au><au>Rish, Adam J.</au><au>Skomo, Alec</au><au>Zhao, Yuxiang</au><au>Drennen, James K.</au><au>Anderson, Carl A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid serum‐free/suspension adaptation: Medium development using a definitive screening design for Chinese hamster ovary cells</atitle><jtitle>Biotechnology progress</jtitle><addtitle>Biotechnol Prog</addtitle><date>2021-07</date><risdate>2021</risdate><volume>37</volume><issue>4</issue><spage>e3154</spage><epage>n/a</epage><pages>e3154-n/a</pages><issn>8756-7938</issn><eissn>1520-6033</eissn><abstract>The biopharmaceutical industry prefers to culture the mammalian cells in suspension with a serum‐free media (SFM) due to improved productivity and process consistency. However, mammalian cells preferentially grow as adherent cells in a complete medium (CM) containing serum. Therefore, cells require adaptation from adherence in CM to suspension culture in SFM. This work proposes an adaptation method that includes media supplementation during the adaption of Chinese hamster ovary cells. As a result, the adaptation was accelerated compared to the traditional repetitive subculturing. Ca2+/Mg2+ supplementation significantly reduced the doubling time compared to the adaptation without supplementation during the adaptation of adherent cells from 100% CM to 75% CM (p < 0.05). Furthermore, a definitive screening design (DSD) was applied to select essential nutrients during the adaptation from 10% CM to 0% CM. The main effects of Ca2+ and Dulbecco's modified essential medium (DMEM) were found significant to both viable cell density and viability at harvest. Additionally, the interaction term between Ca2+ and DMEM was found significant, which highlights the ability of DSD to capture interaction terms. Eventually, the media supplementation method resulted in adaptation SFM in 27 days, compared to the previously reported 66 days. Additionally, the membrane surface integrin expression was found significantly decreased when adherent cells were adapted to suspension. Moreover, the Ca2+/Mg2+ supplementation correlated with faster integrin recovery after trypsinization. However, faster integrin recovery did not contribute to the accelerated cell growth when subculturing from 100% CM to 75% CM.</abstract><cop>Hoboken, USA</cop><pub>John Wiley & Sons, Inc</pub><pmid>33864359</pmid><doi>10.1002/btpr.3154</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-7069-6777</orcidid></addata></record> |
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| subjects | Adaptation Adherent cells Biopharmaceuticals Calcium Calcium ions Cell culture Cell density CHO cells definitive screening design Essential nutrients integrin Magnesium Mammalian cells Mammals Nutrients Ovaries Recovery Screening serum‐free/suspension adaptation Supplements Suspension culture |
| title | Rapid serum‐free/suspension adaptation: Medium development using a definitive screening design for Chinese hamster ovary cells |
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