3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening
A pillar dishe for subculture of 3D cultured cells on hydrogel spots (Matrigel and alginate) have been developed. Cells cultured in 3D in an extracellular matrix (ECM) can retain their intrinsic properties, but cells cultured in 2D lose their intrinsic properties as the cells stick to the bottom of...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | Materials Today Bio |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590006423002533 |
| _version_ | 1797676648036827136 |
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| author | Sang-Yun Lee Hyun Ju Hwang You Jin Song Dayoung Lee Bosung Ku Jason K. Sa Dong Woo Lee |
| author_facet | Sang-Yun Lee Hyun Ju Hwang You Jin Song Dayoung Lee Bosung Ku Jason K. Sa Dong Woo Lee |
| author_sort | Sang-Yun Lee |
| collection | DOAJ |
| description | A pillar dishe for subculture of 3D cultured cells on hydrogel spots (Matrigel and alginate) have been developed. Cells cultured in 3D in an extracellular matrix (ECM) can retain their intrinsic properties, but cells cultured in 2D lose their intrinsic properties as the cells stick to the bottom of the well. Previously, cells and ECM spots were dispensed on a conventional culture dish for 3D cultivation. However, as the spot shape and location depended on user handling, pillars were added to the dish to realize uniform spot shape and stable subculture, supporting 3D cell culture-based high-throughput screening (HTS). Matrigel and alginate were used as ECMs during 6-passage subculture. The growth rate of lung cancer cell (A549) was higher on Matrigel than on alginate. Cancer cell was subcultured in three dimensions in the proposed pillar dish and used for drug screening and differential gene expression analysis. Interestingly, stemness markers, which are unique characteristics of lung cancer cells inducing drug resistance, were upregulated in 3D-subcultured cells compared with those in 2D-subcultured cells. Additionally, the PI3K/Akt/mTOR, VEGFR1/2, and Wnt pathways, which are promising therapeutic targets for lung cancer, were activated, showing high drug sensitivity under 3D-HTS using the 3D-subcultured cells. |
| first_indexed | 2024-03-11T22:31:19Z |
| format | Article |
| id | doaj.art-d5eb530e89bd41338bfdb35ec997da52 |
| institution | Directory Open Access Journal |
| issn | 2590-0064 |
| language | English |
| last_indexed | 2024-03-11T22:31:19Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Materials Today Bio |
| spelling | doaj.art-d5eb530e89bd41338bfdb35ec997da522023-09-23T05:12:02ZengElsevierMaterials Today Bio2590-00642023-12-01231007933D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screeningSang-Yun Lee0Hyun Ju Hwang1You Jin Song2Dayoung Lee3Bosung Ku4Jason K. Sa5Dong Woo Lee6Central R & D Center, Medical & Bio Decision (MBD) Co., Ltd, Suwon, 16229, Republic of Korea; Department of Biomedical Engineering, Gachon University, Seongnam, 13120, Republic of KoreaCentral R & D Center, Medical & Bio Decision (MBD) Co., Ltd, Suwon, 16229, Republic of KoreaDepartment of Biomedical Sciences, Korea University College of Medicine, Seoul, 02841, Republic of KoreaDepartment of Biomedical Sciences, Korea University College of Medicine, Seoul, 02841, Republic of KoreaCentral R & D Center, Medical & Bio Decision (MBD) Co., Ltd, Suwon, 16229, Republic of KoreaDepartment of Biomedical Sciences, Korea University College of Medicine, Seoul, 02841, Republic of Korea; Corresponding author.Department of Biomedical Engineering, Gachon University, Seongnam, 13120, Republic of Korea; Corresponding author.A pillar dishe for subculture of 3D cultured cells on hydrogel spots (Matrigel and alginate) have been developed. Cells cultured in 3D in an extracellular matrix (ECM) can retain their intrinsic properties, but cells cultured in 2D lose their intrinsic properties as the cells stick to the bottom of the well. Previously, cells and ECM spots were dispensed on a conventional culture dish for 3D cultivation. However, as the spot shape and location depended on user handling, pillars were added to the dish to realize uniform spot shape and stable subculture, supporting 3D cell culture-based high-throughput screening (HTS). Matrigel and alginate were used as ECMs during 6-passage subculture. The growth rate of lung cancer cell (A549) was higher on Matrigel than on alginate. Cancer cell was subcultured in three dimensions in the proposed pillar dish and used for drug screening and differential gene expression analysis. Interestingly, stemness markers, which are unique characteristics of lung cancer cells inducing drug resistance, were upregulated in 3D-subcultured cells compared with those in 2D-subcultured cells. Additionally, the PI3K/Akt/mTOR, VEGFR1/2, and Wnt pathways, which are promising therapeutic targets for lung cancer, were activated, showing high drug sensitivity under 3D-HTS using the 3D-subcultured cells.http://www.sciencedirect.com/science/article/pii/S25900064230025333D cell cultureCell culture dishMicropillar and well chipsHigh-throughput screening |
| spellingShingle | Sang-Yun Lee Hyun Ju Hwang You Jin Song Dayoung Lee Bosung Ku Jason K. Sa Dong Woo Lee 3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening Materials Today Bio 3D cell culture Cell culture dish Micropillar and well chips High-throughput screening |
| title | 3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening |
| title_full | 3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening |
| title_fullStr | 3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening |
| title_full_unstemmed | 3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening |
| title_short | 3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening |
| title_sort | 3d cell subculturing pillar dish for pharmacogenetic analysis and high throughput screening |
| topic | 3D cell culture Cell culture dish Micropillar and well chips High-throughput screening |
| url | http://www.sciencedirect.com/science/article/pii/S2590006423002533 |
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