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Microwell bag culture for large-scale production of homogeneous islet-like clusters
Pluripotent stem-cell derived cells can be used for type I diabetes treatment, but we require at least 10 5 –10 6 islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical...
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| Published in: | Scientific reports 2022-03, Vol.12 (1), p.5221-5221, Article 5221 |
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| Main Authors: | , , , , , , |
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| cited_by | cdi_FETCH-LOGICAL-c650t-27f6fd227d3f3f2ac6e8cbee2791610840ef920873338a79c910ec1bb37bf1143 |
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| cites | cdi_FETCH-LOGICAL-c650t-27f6fd227d3f3f2ac6e8cbee2791610840ef920873338a79c910ec1bb37bf1143 |
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| creator | Suenaga, Ryo Konagaya, Shuhei Yamaura, Junji Ito, Ryo Tanaka, Satoshi Ishizaki, Yoichi Toyoda, Taro |
| description | Pluripotent stem-cell derived cells can be used for type I diabetes treatment, but we require at least 10
5
–10
6
islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical use. In this study, we aimed to develop a novel bag culture method for the production of uniform cell clusters on a large scale (10
5
–10
6
clusters). We prepared small-scale culture bags ( |
| doi_str_mv | 10.1038/s41598-022-09124-w |
| format | article |
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5
–10
6
islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical use. In this study, we aimed to develop a novel bag culture method for the production of uniform cell clusters on a large scale (10
5
–10
6
clusters). We prepared small-scale culture bags (< 10
5
clusters) with microwells at the bottom and optimized the conditions for producing uniform-sized clusters in the bag using undifferentiated induced pluripotent stem cells (iPSCs). Subsequently, we verified the suitability of the bag culture method using iPSC-derived pancreatic islet cells (iPICs) and successfully demonstrate the production of 6.5 × 10
5
uniform iPIC clusters using a large-scale bag. In addition, we simplified the pre- and post-process of the culture—a degassing process before cell seeding and a cluster harvesting process. In conclusion, compared with conventional methods, the cluster production method using bags exhibits improved scalability, sterility, and operability for both clinical and research use.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-022-09124-w</identifier><identifier>PMID: 35338209</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/61/2320 ; 631/61/490 ; 631/61/54 ; Cell culture ; Degassing ; Diabetes ; Diabetes mellitus ; Humanities and Social Sciences ; Insulin ; Islet cells ; multidisciplinary ; Permeability ; Pharmaceutical industry ; Pluripotency ; R&D ; Research & development ; Science ; Science (multidisciplinary) ; Stem cells ; Sterility</subject><ispartof>Scientific reports, 2022-03, Vol.12 (1), p.5221-5221, Article 5221</ispartof><rights>The Author(s) 2022</rights><rights>2022. The Author(s).</rights><rights>The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c650t-27f6fd227d3f3f2ac6e8cbee2791610840ef920873338a79c910ec1bb37bf1143</citedby><cites>FETCH-LOGICAL-c650t-27f6fd227d3f3f2ac6e8cbee2791610840ef920873338a79c910ec1bb37bf1143</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2643138567/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2643138567?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793,75126</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35338209$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Suenaga, Ryo</creatorcontrib><creatorcontrib>Konagaya, Shuhei</creatorcontrib><creatorcontrib>Yamaura, Junji</creatorcontrib><creatorcontrib>Ito, Ryo</creatorcontrib><creatorcontrib>Tanaka, Satoshi</creatorcontrib><creatorcontrib>Ishizaki, Yoichi</creatorcontrib><creatorcontrib>Toyoda, Taro</creatorcontrib><title>Microwell bag culture for large-scale production of homogeneous islet-like clusters</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Pluripotent stem-cell derived cells can be used for type I diabetes treatment, but we require at least 10
5
–10
6
islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical use. In this study, we aimed to develop a novel bag culture method for the production of uniform cell clusters on a large scale (10
5
–10
6
clusters). We prepared small-scale culture bags (< 10
5
clusters) with microwells at the bottom and optimized the conditions for producing uniform-sized clusters in the bag using undifferentiated induced pluripotent stem cells (iPSCs). Subsequently, we verified the suitability of the bag culture method using iPSC-derived pancreatic islet cells (iPICs) and successfully demonstrate the production of 6.5 × 10
5
uniform iPIC clusters using a large-scale bag. In addition, we simplified the pre- and post-process of the culture—a degassing process before cell seeding and a cluster harvesting process. In conclusion, compared with conventional methods, the cluster production method using bags exhibits improved scalability, sterility, and operability for both clinical and research use.</description><subject>631/61/2320</subject><subject>631/61/490</subject><subject>631/61/54</subject><subject>Cell culture</subject><subject>Degassing</subject><subject>Diabetes</subject><subject>Diabetes mellitus</subject><subject>Humanities and Social Sciences</subject><subject>Insulin</subject><subject>Islet cells</subject><subject>multidisciplinary</subject><subject>Permeability</subject><subject>Pharmaceutical industry</subject><subject>Pluripotency</subject><subject>R&D</subject><subject>Research & development</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>Stem cells</subject><subject>Sterility</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNp9kk1vFSEUhonR2ObaP-DCTOLGDcrXzMDGxDR-NKlxoa4JMIcpV-5whZne-O-ld2ptXcgGwnl5OB8vQs8peU0Jl2-KoK2SmDCGiaJM4MMjdMqIaDHjjD2-dz5BZ6VsSV0tU4Kqp-iEt5xLRtQp-vo5uJwOEGNjzdi4Jc5Lhsan3ESTR8DFmQjNPqdhcXNIU5N8c5V2aYQJ0lKaUCLMOIYf0Li4lBlyeYaeeBMLnN3uG_T9w_tv55_w5ZePF-fvLrHrWjJj1vvOD4z1A_fcM-M6kM4CsF7RjhIpCHjFiOx5Tdb0yilKwFFreW89pYJv0MXKHZLZ6n0OO5N_6WSCPl6kPGqT5-AiaC-pYkYQKzgVYhDKAvUeBmUNl-BsZb1dWfvF7mBwMM3ZxAfQh5EpXOkxXWup2q7jsgJe3QJy-rlAmfUuFFf7ao590qwTghDe1Wo26OU_0m1a8lRbdaPilMu2yjaIrao6n1Iy-LtkKNE3FtCrBXS1gD5aQB_qoxf3y7h78mfgVcBXQamhaYT89-__YH8DTbO9xg</recordid><startdate>20220325</startdate><enddate>20220325</enddate><creator>Suenaga, Ryo</creator><creator>Konagaya, Shuhei</creator><creator>Yamaura, Junji</creator><creator>Ito, Ryo</creator><creator>Tanaka, Satoshi</creator><creator>Ishizaki, Yoichi</creator><creator>Toyoda, Taro</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><general>Nature Portfolio</general><scope>C6C</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20220325</creationdate><title>Microwell bag culture for large-scale production of homogeneous islet-like clusters</title><author>Suenaga, Ryo ; Konagaya, Shuhei ; Yamaura, Junji ; Ito, Ryo ; Tanaka, Satoshi ; Ishizaki, Yoichi ; Toyoda, Taro</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c650t-27f6fd227d3f3f2ac6e8cbee2791610840ef920873338a79c910ec1bb37bf1143</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>631/61/2320</topic><topic>631/61/490</topic><topic>631/61/54</topic><topic>Cell culture</topic><topic>Degassing</topic><topic>Diabetes</topic><topic>Diabetes mellitus</topic><topic>Humanities and Social Sciences</topic><topic>Insulin</topic><topic>Islet cells</topic><topic>multidisciplinary</topic><topic>Permeability</topic><topic>Pharmaceutical industry</topic><topic>Pluripotency</topic><topic>R&D</topic><topic>Research & development</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Stem cells</topic><topic>Sterility</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Suenaga, Ryo</creatorcontrib><creatorcontrib>Konagaya, Shuhei</creatorcontrib><creatorcontrib>Yamaura, Junji</creatorcontrib><creatorcontrib>Ito, Ryo</creatorcontrib><creatorcontrib>Tanaka, Satoshi</creatorcontrib><creatorcontrib>Ishizaki, Yoichi</creatorcontrib><creatorcontrib>Toyoda, Taro</creatorcontrib><collection>SpringerOpen</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>ProQuest Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science 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>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>ProQuest Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection (Proquest) (PQ_SDU_P3)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biological Sciences</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest Science Journals</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database (Proquest) (PQ_SDU_P3)</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 Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Suenaga, Ryo</au><au>Konagaya, Shuhei</au><au>Yamaura, Junji</au><au>Ito, Ryo</au><au>Tanaka, Satoshi</au><au>Ishizaki, Yoichi</au><au>Toyoda, Taro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Microwell bag culture for large-scale production of homogeneous islet-like clusters</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2022-03-25</date><risdate>2022</risdate><volume>12</volume><issue>1</issue><spage>5221</spage><epage>5221</epage><pages>5221-5221</pages><artnum>5221</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Pluripotent stem-cell derived cells can be used for type I diabetes treatment, but we require at least 10
5
–10
6
islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical use. In this study, we aimed to develop a novel bag culture method for the production of uniform cell clusters on a large scale (10
5
–10
6
clusters). We prepared small-scale culture bags (< 10
5
clusters) with microwells at the bottom and optimized the conditions for producing uniform-sized clusters in the bag using undifferentiated induced pluripotent stem cells (iPSCs). Subsequently, we verified the suitability of the bag culture method using iPSC-derived pancreatic islet cells (iPICs) and successfully demonstrate the production of 6.5 × 10
5
uniform iPIC clusters using a large-scale bag. In addition, we simplified the pre- and post-process of the culture—a degassing process before cell seeding and a cluster harvesting process. In conclusion, compared with conventional methods, the cluster production method using bags exhibits improved scalability, sterility, and operability for both clinical and research use.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>35338209</pmid><doi>10.1038/s41598-022-09124-w</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| language | eng |
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| subjects | 631/61/2320 631/61/490 631/61/54 Cell culture Degassing Diabetes Diabetes mellitus Humanities and Social Sciences Insulin Islet cells multidisciplinary Permeability Pharmaceutical industry Pluripotency R&D Research & development Science Science (multidisciplinary) Stem cells Sterility |
| title | Microwell bag culture for large-scale production of homogeneous islet-like clusters |
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