Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models
Abstract The epithelial and mesenchymal features of colorectal adenocarcinoma (CRAC) cell lines were compared in two-dimensional (2D) and three-dimensional (3D) cultures. In 2D cultures, the three CRAC cell lines exhibited epithelial characteristics with high E-cadherin and low vimentin levels, wher...
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Nature Portfolio
2025-01-01
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| Online Access: | https://doi.org/10.1038/s41598-025-87660-x |
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| author | Seiichi Shinji Yutaro Ogawa Takeshi Yamada Akihisa Matsuda Kay Uehara Yasuyuki Yokoyama Goro Takahashi Takuma Iwai Toshimitsu Miyasaka Shintaro Kanaka Koki Hayashi Yuuki Shichi Masakazu Fujiwara Kimimasa Takahashi Tomio Arai Toshiyuki Ishiwata Hiroshi Yoshida |
| author_facet | Seiichi Shinji Yutaro Ogawa Takeshi Yamada Akihisa Matsuda Kay Uehara Yasuyuki Yokoyama Goro Takahashi Takuma Iwai Toshimitsu Miyasaka Shintaro Kanaka Koki Hayashi Yuuki Shichi Masakazu Fujiwara Kimimasa Takahashi Tomio Arai Toshiyuki Ishiwata Hiroshi Yoshida |
| author_sort | Seiichi Shinji |
| collection | DOAJ |
| description | Abstract The epithelial and mesenchymal features of colorectal adenocarcinoma (CRAC) cell lines were compared in two-dimensional (2D) and three-dimensional (3D) cultures. In 2D cultures, the three CRAC cell lines exhibited epithelial characteristics with high E-cadherin and low vimentin levels, whereas two exhibited mesenchymal traits with opposite expression patterns. In 3D cultures using low-attachment plates, mesenchymal cells from 2D cultures showed reduced vimentin mRNA levels. Morphologically, the five CRAC cell lines appeared similarly shaped in 2D culture but formed different structures in 3D culture. Epithelial DLD-1 and mesenchymal COLO-320 cells produced large granular spheres, whereas epithelial HCT-15 cells formed small solid spheres. Tubular structures were observed in epithelial CACO-2 and mesenchymal SW480 spheres. Desmosome-like structures developed in epithelial CRAC cells, whereas entosis was observed in CACO-2, HCT-15, and SW480 cells. The Ki-67-positive proliferating cell count varied in 2D and 3D cultures of epithelial cells but remained high and unchanged in mesenchymal cells. These findings suggest that while CRAC cells display distinct epithelial and mesenchymal properties in 2D cultures, they form diverse 3D structures, irrespective of these traits. |
| format | Article |
| id | doaj-art-4c07a1a0d3c945b4b5e308831ddc7d42 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-4c07a1a0d3c945b4b5e308831ddc7d422025-01-26T12:31:15ZengNature PortfolioScientific Reports2045-23222025-01-0115111110.1038/s41598-025-87660-xMorphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture modelsSeiichi Shinji0Yutaro Ogawa1Takeshi Yamada2Akihisa Matsuda3Kay Uehara4Yasuyuki Yokoyama5Goro Takahashi6Takuma Iwai7Toshimitsu Miyasaka8Shintaro Kanaka9Koki Hayashi10Yuuki Shichi11Masakazu Fujiwara12Kimimasa Takahashi13Tomio Arai14Toshiyuki Ishiwata15Hiroshi Yoshida16Department of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDepartment of Gastroenterological Surgery, Nippon Medical SchoolDivision of Aging and Carcinogenesis, Research Team for Geriatric Pathology, Tokyo Metropolitan Institute for Geriatrics and GerontologyDivision of Aging and Carcinogenesis, Research Team for Geriatric Pathology, Tokyo Metropolitan Institute for Geriatrics and GerontologyDepartment of Veterinary Pathology, School of Veterinary Medicine, Nippon Veterinary and Life Science UniversityDepartment of Pathology, Tokyo Metropolitan Institute for Geriatrics and GerontologyDivision of Aging and Carcinogenesis, Research Team for Geriatric Pathology, Tokyo Metropolitan Institute for Geriatrics and GerontologyDepartment of Gastroenterological Surgery, Nippon Medical SchoolAbstract The epithelial and mesenchymal features of colorectal adenocarcinoma (CRAC) cell lines were compared in two-dimensional (2D) and three-dimensional (3D) cultures. In 2D cultures, the three CRAC cell lines exhibited epithelial characteristics with high E-cadherin and low vimentin levels, whereas two exhibited mesenchymal traits with opposite expression patterns. In 3D cultures using low-attachment plates, mesenchymal cells from 2D cultures showed reduced vimentin mRNA levels. Morphologically, the five CRAC cell lines appeared similarly shaped in 2D culture but formed different structures in 3D culture. Epithelial DLD-1 and mesenchymal COLO-320 cells produced large granular spheres, whereas epithelial HCT-15 cells formed small solid spheres. Tubular structures were observed in epithelial CACO-2 and mesenchymal SW480 spheres. Desmosome-like structures developed in epithelial CRAC cells, whereas entosis was observed in CACO-2, HCT-15, and SW480 cells. The Ki-67-positive proliferating cell count varied in 2D and 3D cultures of epithelial cells but remained high and unchanged in mesenchymal cells. These findings suggest that while CRAC cells display distinct epithelial and mesenchymal properties in 2D cultures, they form diverse 3D structures, irrespective of these traits.https://doi.org/10.1038/s41598-025-87660-xColorectal adenocarcinoma2-dimensional culture3-dimensional cultureSphereScanning electron microscopyTransmission electron microscopy |
| spellingShingle | Seiichi Shinji Yutaro Ogawa Takeshi Yamada Akihisa Matsuda Kay Uehara Yasuyuki Yokoyama Goro Takahashi Takuma Iwai Toshimitsu Miyasaka Shintaro Kanaka Koki Hayashi Yuuki Shichi Masakazu Fujiwara Kimimasa Takahashi Tomio Arai Toshiyuki Ishiwata Hiroshi Yoshida Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models Scientific Reports Colorectal adenocarcinoma 2-dimensional culture 3-dimensional culture Sphere Scanning electron microscopy Transmission electron microscopy |
| title | Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models |
| title_full | Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models |
| title_fullStr | Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models |
| title_full_unstemmed | Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models |
| title_short | Morphological and functional analysis of colorectal cancer cell lines in 2D and 3D culture models |
| title_sort | morphological and functional analysis of colorectal cancer cell lines in 2d and 3d culture models |
| topic | Colorectal adenocarcinoma 2-dimensional culture 3-dimensional culture Sphere Scanning electron microscopy Transmission electron microscopy |
| url | https://doi.org/10.1038/s41598-025-87660-x |
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