Novel and efficient method for culturing patient-derived gastric cancer stem cells.
Wnt
extracellular matrix
gastric cancer
spheroid
stem cell
Journal
Cancer science
ISSN: 1349-7006
Titre abrégé: Cancer Sci
Pays: England
ID NLM: 101168776
Informations de publication
Date de publication:
Aug 2023
Aug 2023
Historique:
revised:
16
04
2023
received:
06
12
2022
accepted:
21
04
2023
medline:
3
8
2023
pubmed:
20
5
2023
entrez:
20
5
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 37208931
doi: 10.1111/cas.15840
pmc: PMC10394150
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3259-3269Subventions
Organisme : Institute for Advancement of Clinical and Translational Science, Kyoto University Hospital
Organisme : Japan Agency for Medical Research and Development
ID : ck0106195h
Organisme : Japan Science and Technology Agency
ID : ST261001TT
Organisme : Japan Society for the Promotion of Science
ID : JP18H02639
Organisme : Japan Society for the Promotion of Science
ID : JP21K06948
Organisme : Japan Society for the Promotion of Science
ID : JP22K07187
Organisme : Kyo Diagnostics K.K.
Organisme : Kyoto University Office of Society-Academia Collaboration for Innovation
Organisme : SCREEN Holdings Co., Ltd.
Informations de copyright
© 2023 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.
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