Novel and efficient method for culturing patient‐derived gastric cancer stem cells
Experimental techniques for patient‐derived cancer stem‐cell organoids/spheroids can be powerful diagnostic tools for personalized chemotherapy. However, establishing their cultures from gastric cancer remains challenging due to low culture efficiency and cumbersome methods. To propagate gastric can...
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| Veröffentlicht in: | Cancer science 2023-08, Vol.114 (8), p.3259-3269 |
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| creator | Morimoto, Tomonori Takemura, Yukitoshi Miura, Takemitsu Yamamoto, Takehito Kakizaki, Fumihiko An, Hideo Maekawa, Hisatsugu Yamaura, Tadayoshi Kawada, Kenji Sakai, Yoshiharu Yuba, Yoshiaki Terajima, Hiroaki Obama, Kazutaka Taketo, Makoto Mark Miyoshi, Hiroyuki |
| description | Experimental techniques for patient‐derived cancer stem‐cell organoids/spheroids can be powerful diagnostic tools for personalized chemotherapy. However, establishing their cultures from gastric cancer remains challenging due to low culture efficiency and cumbersome methods. To propagate gastric cancer cells as highly proliferative stem‐cell spheroids in vitro, we initially used a similar method to that for colorectal cancer stem cells, which, unfortunately, resulted in a low success rate (25%, 18 of 71 cases). We scrutinized the protocol and found that the unsuccessful cases were largely caused by the paucity of cancer stem cells in the sampled tissues as well as insufficient culture media. To overcome these obstacles, we extensively revised our sample collection protocol and culture conditions. We then investigated the following second cohort and, consequently, achieved a significantly higher success rate (88%, 29 of 33 cases). One of the key improvements included new sampling procedures for tumor tissues from wider and deeper areas of gastric cancer specimens, which allowed securing cancer stem cells more reproducibly. Additionally, we embedded tumor epithelial pieces separately in both Matrigel and collagen type‐I as their preference to the extracellular matrix was different depending on the tumors. We also added a low concentration of Wnt ligands to the culture, which helped the growth of occasional Wnt‐responsive gastric cancer stem‐cell spheroids without allowing proliferation of the normal gastric epithelial stem cells. This newly improved spheroid culture method may facilitate further studies, including personalized drug‐sensitivity tests prior to drug therapy.
We developed a simple and efficient method to propagate patient‐derived gastric cancer stem‐cell spheroids by improving our conventional sample collection protocol and culture conditions. These spheroids can be utilized to investigate new molecular targeted therapies and their companion diagnostics for patient selection. |
| doi_str_mv | 10.1111/cas.15840 |
| format | Article |
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We developed a simple and efficient method to propagate patient‐derived gastric cancer stem‐cell spheroids by improving our conventional sample collection protocol and culture conditions. These spheroids can be utilized to investigate new molecular targeted therapies and their companion diagnostics for patient selection.</description><identifier>ISSN: 1347-9032</identifier><identifier>EISSN: 1349-7006</identifier><identifier>DOI: 10.1111/cas.15840</identifier><identifier>PMID: 37208931</identifier><language>eng</language><publisher>England: John Wiley & Sons, Inc</publisher><subject>Cancer therapies ; Cell culture ; Cell growth ; Cell proliferation ; Chemotherapy ; Collagen ; Colorectal cancer ; Colorectal carcinoma ; Culture media ; Drug therapy ; Epidermal growth factor ; Extracellular matrix ; Gastric cancer ; Genes ; Genomes ; Genomics ; Medical research ; Mutation ; Organoids ; Original ; ORIGINAL ARTICLES ; Patients ; Polymerization ; Software ; spheroid ; Spheroids ; stem cell ; Stem cells ; Success ; Tumors ; Wnt ; Wnt protein</subject><ispartof>Cancer science, 2023-08, Vol.114 (8), p.3259-3269</ispartof><rights>2023 The Authors. published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.</rights><rights>2023 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.</rights><rights>2023. This work is published under http://creativecommons.org/licenses/by-nc/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><cites>FETCH-LOGICAL-c4280-21987e2b3ee2ecb7a4981189fbf5f3c2a47dfb8bc08cda8c8812f17d2972a1d83</cites><orcidid>0000-0001-7400-0714 ; 0000-0003-0535-0889 ; 0000-0003-3066-6658 ; 0000-0003-2017-6700 ; 0000-0002-2236-7124 ; 0000-0003-0019-1313 ; 0000-0003-4336-6937 ; 0000-0002-9032-4505 ; 0000-0003-4565-529X ; 0000-0002-0200-414X ; 0009-0004-4762-7617 ; 0000-0001-7938-6345 ; 0000-0003-2924-6701 ; 0000-0001-9864-4186</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC10394150/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC10394150/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,1417,11562,27924,27925,45574,45575,46052,46476,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37208931$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Morimoto, Tomonori</creatorcontrib><creatorcontrib>Takemura, Yukitoshi</creatorcontrib><creatorcontrib>Miura, Takemitsu</creatorcontrib><creatorcontrib>Yamamoto, Takehito</creatorcontrib><creatorcontrib>Kakizaki, Fumihiko</creatorcontrib><creatorcontrib>An, Hideo</creatorcontrib><creatorcontrib>Maekawa, Hisatsugu</creatorcontrib><creatorcontrib>Yamaura, Tadayoshi</creatorcontrib><creatorcontrib>Kawada, Kenji</creatorcontrib><creatorcontrib>Sakai, Yoshiharu</creatorcontrib><creatorcontrib>Yuba, Yoshiaki</creatorcontrib><creatorcontrib>Terajima, Hiroaki</creatorcontrib><creatorcontrib>Obama, Kazutaka</creatorcontrib><creatorcontrib>Taketo, Makoto Mark</creatorcontrib><creatorcontrib>Miyoshi, Hiroyuki</creatorcontrib><title>Novel and efficient method for culturing patient‐derived gastric cancer stem cells</title><title>Cancer science</title><addtitle>Cancer Sci</addtitle><description>Experimental techniques for patient‐derived cancer stem‐cell organoids/spheroids can be powerful diagnostic tools for personalized chemotherapy. However, establishing their cultures from gastric cancer remains challenging due to low culture efficiency and cumbersome methods. To propagate gastric cancer cells as highly proliferative stem‐cell spheroids in vitro, we initially used a similar method to that for colorectal cancer stem cells, which, unfortunately, resulted in a low success rate (25%, 18 of 71 cases). We scrutinized the protocol and found that the unsuccessful cases were largely caused by the paucity of cancer stem cells in the sampled tissues as well as insufficient culture media. To overcome these obstacles, we extensively revised our sample collection protocol and culture conditions. We then investigated the following second cohort and, consequently, achieved a significantly higher success rate (88%, 29 of 33 cases). One of the key improvements included new sampling procedures for tumor tissues from wider and deeper areas of gastric cancer specimens, which allowed securing cancer stem cells more reproducibly. Additionally, we embedded tumor epithelial pieces separately in both Matrigel and collagen type‐I as their preference to the extracellular matrix was different depending on the tumors. We also added a low concentration of Wnt ligands to the culture, which helped the growth of occasional Wnt‐responsive gastric cancer stem‐cell spheroids without allowing proliferation of the normal gastric epithelial stem cells. This newly improved spheroid culture method may facilitate further studies, including personalized drug‐sensitivity tests prior to drug therapy.
We developed a simple and efficient method to propagate patient‐derived gastric cancer stem‐cell spheroids by improving our conventional sample collection protocol and culture conditions. These spheroids can be utilized to investigate new molecular targeted therapies and their companion diagnostics for patient selection.</description><subject>Cancer therapies</subject><subject>Cell culture</subject><subject>Cell growth</subject><subject>Cell proliferation</subject><subject>Chemotherapy</subject><subject>Collagen</subject><subject>Colorectal cancer</subject><subject>Colorectal carcinoma</subject><subject>Culture media</subject><subject>Drug therapy</subject><subject>Epidermal growth factor</subject><subject>Extracellular matrix</subject><subject>Gastric cancer</subject><subject>Genes</subject><subject>Genomes</subject><subject>Genomics</subject><subject>Medical research</subject><subject>Mutation</subject><subject>Organoids</subject><subject>Original</subject><subject>ORIGINAL ARTICLES</subject><subject>Patients</subject><subject>Polymerization</subject><subject>Software</subject><subject>spheroid</subject><subject>Spheroids</subject><subject>stem cell</subject><subject>Stem cells</subject><subject>Success</subject><subject>Tumors</subject><subject>Wnt</subject><subject>Wnt protein</subject><issn>1347-9032</issn><issn>1349-7006</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kc9KXDEUh4O0VGu78AVKoBu7uJp_M0lWIkNtC1IX2nXITU7GyL030-TeEXc-Qp_RJ2nGsdIKzeYEzsfH7_BD6ICSI1rfsbPliM6UIDtoj3KhG0nI_NXjXzaacLaL3pZyQwifCy3eoF0uGVGa0z109T2tocN28BhCiC7CMOIexuvkcUgZu6kbpxyHJV7ZcbN8uP_lIcc1eLy0ZczRYWcHBxmXEXrsoOvKO_Q62K7A-6e5j36cfb5afG3OL758W5yeN04wRRpGtZLAWg7AwLXSCq0oVTq0YRa4Y1ZIH1rVOqKct8opRVmg0jMtmaVe8X10svWuprYH72q8bDuzyrG3-c4kG82_myFem2VaG0q4FnRGquHwyZDTzwnKaPpYNjfYAdJUDFN0LmdqTnhFP75Ab9KUh3pfpYQQXAjNKvVpS7mcSskQntNQYjZlmVqWeSyrsh_-jv9M_mmnAsdb4DZ2cPd_k1mcXm6VvwFHhaBL</recordid><startdate>202308</startdate><enddate>202308</enddate><creator>Morimoto, Tomonori</creator><creator>Takemura, Yukitoshi</creator><creator>Miura, Takemitsu</creator><creator>Yamamoto, Takehito</creator><creator>Kakizaki, Fumihiko</creator><creator>An, Hideo</creator><creator>Maekawa, Hisatsugu</creator><creator>Yamaura, Tadayoshi</creator><creator>Kawada, Kenji</creator><creator>Sakai, Yoshiharu</creator><creator>Yuba, Yoshiaki</creator><creator>Terajima, Hiroaki</creator><creator>Obama, Kazutaka</creator><creator>Taketo, Makoto Mark</creator><creator>Miyoshi, Hiroyuki</creator><general>John Wiley & Sons, Inc</general><general>John Wiley and Sons Inc</general><scope>24P</scope><scope>WIN</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FE</scope><scope>8FH</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>GNUQQ</scope><scope>HCIFZ</scope><scope>LK8</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-7400-0714</orcidid><orcidid>https://orcid.org/0000-0003-0535-0889</orcidid><orcidid>https://orcid.org/0000-0003-3066-6658</orcidid><orcidid>https://orcid.org/0000-0003-2017-6700</orcidid><orcidid>https://orcid.org/0000-0002-2236-7124</orcidid><orcidid>https://orcid.org/0000-0003-0019-1313</orcidid><orcidid>https://orcid.org/0000-0003-4336-6937</orcidid><orcidid>https://orcid.org/0000-0002-9032-4505</orcidid><orcidid>https://orcid.org/0000-0003-4565-529X</orcidid><orcidid>https://orcid.org/0000-0002-0200-414X</orcidid><orcidid>https://orcid.org/0009-0004-4762-7617</orcidid><orcidid>https://orcid.org/0000-0001-7938-6345</orcidid><orcidid>https://orcid.org/0000-0003-2924-6701</orcidid><orcidid>https://orcid.org/0000-0001-9864-4186</orcidid></search><sort><creationdate>202308</creationdate><title>Novel and efficient method for culturing patient‐derived gastric cancer stem cells</title><author>Morimoto, Tomonori ; 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However, establishing their cultures from gastric cancer remains challenging due to low culture efficiency and cumbersome methods. To propagate gastric cancer cells as highly proliferative stem‐cell spheroids in vitro, we initially used a similar method to that for colorectal cancer stem cells, which, unfortunately, resulted in a low success rate (25%, 18 of 71 cases). We scrutinized the protocol and found that the unsuccessful cases were largely caused by the paucity of cancer stem cells in the sampled tissues as well as insufficient culture media. To overcome these obstacles, we extensively revised our sample collection protocol and culture conditions. We then investigated the following second cohort and, consequently, achieved a significantly higher success rate (88%, 29 of 33 cases). One of the key improvements included new sampling procedures for tumor tissues from wider and deeper areas of gastric cancer specimens, which allowed securing cancer stem cells more reproducibly. Additionally, we embedded tumor epithelial pieces separately in both Matrigel and collagen type‐I as their preference to the extracellular matrix was different depending on the tumors. We also added a low concentration of Wnt ligands to the culture, which helped the growth of occasional Wnt‐responsive gastric cancer stem‐cell spheroids without allowing proliferation of the normal gastric epithelial stem cells. This newly improved spheroid culture method may facilitate further studies, including personalized drug‐sensitivity tests prior to drug therapy.
We developed a simple and efficient method to propagate patient‐derived gastric cancer stem‐cell spheroids by improving our conventional sample collection protocol and culture conditions. These spheroids can be utilized to investigate new molecular targeted therapies and their companion diagnostics for patient selection.</abstract><cop>England</cop><pub>John Wiley & Sons, Inc</pub><pmid>37208931</pmid><doi>10.1111/cas.15840</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-7400-0714</orcidid><orcidid>https://orcid.org/0000-0003-0535-0889</orcidid><orcidid>https://orcid.org/0000-0003-3066-6658</orcidid><orcidid>https://orcid.org/0000-0003-2017-6700</orcidid><orcidid>https://orcid.org/0000-0002-2236-7124</orcidid><orcidid>https://orcid.org/0000-0003-0019-1313</orcidid><orcidid>https://orcid.org/0000-0003-4336-6937</orcidid><orcidid>https://orcid.org/0000-0002-9032-4505</orcidid><orcidid>https://orcid.org/0000-0003-4565-529X</orcidid><orcidid>https://orcid.org/0000-0002-0200-414X</orcidid><orcidid>https://orcid.org/0009-0004-4762-7617</orcidid><orcidid>https://orcid.org/0000-0001-7938-6345</orcidid><orcidid>https://orcid.org/0000-0003-2924-6701</orcidid><orcidid>https://orcid.org/0000-0001-9864-4186</orcidid><oa>free_for_read</oa></addata></record> |
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| source | Wiley Online Library - AutoHoldings Journals; DOAJ Directory of Open Access Journals; Wiley Online Library Open Access; PubMed |
| subjects | Cancer therapies Cell culture Cell growth Cell proliferation Chemotherapy Collagen Colorectal cancer Colorectal carcinoma Culture media Drug therapy Epidermal growth factor Extracellular matrix Gastric cancer Genes Genomes Genomics Medical research Mutation Organoids Original ORIGINAL ARTICLES Patients Polymerization Software spheroid Spheroids stem cell Stem cells Success Tumors Wnt Wnt protein |
| title | Novel and efficient method for culturing patient‐derived gastric cancer stem cells |
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