Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties
Spheroids are expected to aid the establishment of an in vitro-based cell culture system that can realistically reproduce cellular dynamics in vivo. We developed a fluoropolymer scaffold with an extracellular matrix (ECM) dot array and confirmed the possibility of mass-producing spheroids with unifo...
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| Format: | Article |
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MDPI AG
2023-01-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/2/278 |
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| author | Hidetaka Togo Kento Terada Akira Ujitsugu Yudai Hirose Hiroki Takeuchi Masanobu Kusunoki |
| author_facet | Hidetaka Togo Kento Terada Akira Ujitsugu Yudai Hirose Hiroki Takeuchi Masanobu Kusunoki |
| author_sort | Hidetaka Togo |
| collection | DOAJ |
| description | Spheroids are expected to aid the establishment of an in vitro-based cell culture system that can realistically reproduce cellular dynamics in vivo. We developed a fluoropolymer scaffold with an extracellular matrix (ECM) dot array and confirmed the possibility of mass-producing spheroids with uniform dimensions. Controlling the quality of ECM dots is important as it ensures spheroid uniformity, but issues such as pattern deviation and ECM drying persist in the conventional microstamping method. In this study, these problems were overcome via ECM dot printing using a resin mask with dot-patterned holes. For dot diameters of φ 300 μm, 400 μm, and 600 μm, the average spheroid diameters of human iPS cells (hiPSCs) were φ 260.8 μm, 292.4 μm, and 330.7 μm, respectively. The standard deviation when each average was normalized to 100 was 14.1%. A high throughput of 89.9% for colony formation rate to the number of dots and 89.3% for spheroid collection rate was achieved. The cells proliferated on ECM dots, and the colonies could be naturally detached from the scaffold without the use of enzymes, so there was almost no stimulation of the cells. Thus, the undifferentiated nature of hiPSCs was maintained until day 4. Therefore, this method is expected to be useful in drug discovery and regenerative medicine. |
| first_indexed | 2024-03-09T13:13:03Z |
| format | Article |
| id | doaj.art-e6e37aa1e02f409b9102b0fcfdba0df8 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T13:13:03Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-e6e37aa1e02f409b9102b0fcfdba0df82023-11-30T21:40:13ZengMDPI AGCells2073-44092023-01-0112227810.3390/cells12020278Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell PropertiesHidetaka Togo0Kento Terada1Akira Ujitsugu2Yudai Hirose3Hiroki Takeuchi4Masanobu Kusunoki5Graduate School of Biology-Oriented-Science and Technology, Kindai University, 930 Nishimitani, Kinokawa 649-6493, Wakayama, JapanDepartment of Obstetrics and Gynecology, Graduate School of Medicine, Mie University, 2-174 Edobashi, Tsu 514-8507, Mie, JapanFaculty of Biology-Oriented-Science and Technology, Kindai University, 930 Nishimitani, Kinokawa 649-6493, Wakayama, JapanGraduate School of Biology-Oriented-Science and Technology, Kindai University, 930 Nishimitani, Kinokawa 649-6493, Wakayama, JapanDepartment of Obstetrics and Gynecology, Graduate School of Medicine, Mie University, 2-174 Edobashi, Tsu 514-8507, Mie, JapanGraduate School of Biology-Oriented-Science and Technology, Kindai University, 930 Nishimitani, Kinokawa 649-6493, Wakayama, JapanSpheroids are expected to aid the establishment of an in vitro-based cell culture system that can realistically reproduce cellular dynamics in vivo. We developed a fluoropolymer scaffold with an extracellular matrix (ECM) dot array and confirmed the possibility of mass-producing spheroids with uniform dimensions. Controlling the quality of ECM dots is important as it ensures spheroid uniformity, but issues such as pattern deviation and ECM drying persist in the conventional microstamping method. In this study, these problems were overcome via ECM dot printing using a resin mask with dot-patterned holes. For dot diameters of φ 300 μm, 400 μm, and 600 μm, the average spheroid diameters of human iPS cells (hiPSCs) were φ 260.8 μm, 292.4 μm, and 330.7 μm, respectively. The standard deviation when each average was normalized to 100 was 14.1%. A high throughput of 89.9% for colony formation rate to the number of dots and 89.3% for spheroid collection rate was achieved. The cells proliferated on ECM dots, and the colonies could be naturally detached from the scaffold without the use of enzymes, so there was almost no stimulation of the cells. Thus, the undifferentiated nature of hiPSCs was maintained until day 4. Therefore, this method is expected to be useful in drug discovery and regenerative medicine.https://www.mdpi.com/2073-4409/12/2/278spheroidorganoiduniform dimensionscaffoldiPSundifferentiated nature |
| spellingShingle | Hidetaka Togo Kento Terada Akira Ujitsugu Yudai Hirose Hiroki Takeuchi Masanobu Kusunoki Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties Cells spheroid organoid uniform dimension scaffold iPS undifferentiated nature |
| title | Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties |
| title_full | Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties |
| title_fullStr | Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties |
| title_full_unstemmed | Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties |
| title_short | Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties |
| title_sort | fabrication scaffold with high dimensional control for spheroids with undifferentiated ips cell properties |
| topic | spheroid organoid uniform dimension scaffold iPS undifferentiated nature |
| url | https://www.mdpi.com/2073-4409/12/2/278 |
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