Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages
Astrocytes, a type of glial cell in the brain, are thought to be functionally and morphologically diverse cells that regulate brain homeostasis. Cell immortalization is a promising technique for the propagation of primary human astrocytes. The immortalized cells retain their astrocytic marker mRNA e...
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MDPI AG
2023-03-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/10/3/349 |
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| author | Fumiya Tao Keita Kitamura Sanshiro Hanada Kazuyuki Sugimoto Tomomi Furihata Nobuhiko Kojima |
| author_facet | Fumiya Tao Keita Kitamura Sanshiro Hanada Kazuyuki Sugimoto Tomomi Furihata Nobuhiko Kojima |
| author_sort | Fumiya Tao |
| collection | DOAJ |
| description | Astrocytes, a type of glial cell in the brain, are thought to be functionally and morphologically diverse cells that regulate brain homeostasis. Cell immortalization is a promising technique for the propagation of primary human astrocytes. The immortalized cells retain their astrocytic marker mRNA expression at lower levels than the primary cells. Therefore, improvement of the differentiation status is required. The use of a 3D formation technique to mimic structural tissue is a good strategy for reflecting physiological cell–cell interactions. Previously, we developed a spheroid formation method using highly viscous methyl cellulose (MC) medium. In this study, we applied this formation method to the well-established immortalized human astrocyte cell line HASTR/ci35. Stable HASTR/ci35 spheroids were successfully formed in MC medium, and laminin deposition was detected inside of the spheroids. Their functional markers were enhanced compared to conventional spheroids formed in U-bottom plates. The inflammatory response was moderately sensitive, and the ability to support neurite growth was confirmed. The HASTR/ci35 spheroid in the MC medium demonstrated the differentiation phenotype and could serve as a potent in vitro model for matured astrocytes. |
| first_indexed | 2024-03-11T06:55:58Z |
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| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-11T06:55:58Z |
| publishDate | 2023-03-01 |
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| series | Bioengineering |
| spelling | doaj.art-e5cabafd54c841b793846e6e0df6a6c72023-11-17T09:40:03ZengMDPI AGBioengineering2306-53542023-03-0110334910.3390/bioengineering10030349Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional AdvantagesFumiya Tao0Keita Kitamura1Sanshiro Hanada2Kazuyuki Sugimoto3Tomomi Furihata4Nobuhiko Kojima5Department of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama, Kanagawa 236-0027, JapanLaboratory of Clinical Pharmacy and Experimental Therapeutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0355, JapanDepartment of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama, Kanagawa 236-0027, JapanYokogawa Electric Corp., 2-3, Hokuyodai, Kanazawa, Ishikawa 920-0177, JapanLaboratory of Clinical Pharmacy and Experimental Therapeutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0355, JapanDepartment of Life and Environmental System Science, Graduate School of Nanobioscience, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama, Kanagawa 236-0027, JapanAstrocytes, a type of glial cell in the brain, are thought to be functionally and morphologically diverse cells that regulate brain homeostasis. Cell immortalization is a promising technique for the propagation of primary human astrocytes. The immortalized cells retain their astrocytic marker mRNA expression at lower levels than the primary cells. Therefore, improvement of the differentiation status is required. The use of a 3D formation technique to mimic structural tissue is a good strategy for reflecting physiological cell–cell interactions. Previously, we developed a spheroid formation method using highly viscous methyl cellulose (MC) medium. In this study, we applied this formation method to the well-established immortalized human astrocyte cell line HASTR/ci35. Stable HASTR/ci35 spheroids were successfully formed in MC medium, and laminin deposition was detected inside of the spheroids. Their functional markers were enhanced compared to conventional spheroids formed in U-bottom plates. The inflammatory response was moderately sensitive, and the ability to support neurite growth was confirmed. The HASTR/ci35 spheroid in the MC medium demonstrated the differentiation phenotype and could serve as a potent in vitro model for matured astrocytes.https://www.mdpi.com/2306-5354/10/3/349astrocytescell immortalizationdifferentiation statusspheroid formation method |
| spellingShingle | Fumiya Tao Keita Kitamura Sanshiro Hanada Kazuyuki Sugimoto Tomomi Furihata Nobuhiko Kojima Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages Bioengineering astrocytes cell immortalization differentiation status spheroid formation method |
| title | Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages |
| title_full | Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages |
| title_fullStr | Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages |
| title_full_unstemmed | Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages |
| title_short | Rapid and Stable Formation Method of Human Astrocyte Spheroid in a High Viscous Methylcellulose Medium and Its Functional Advantages |
| title_sort | rapid and stable formation method of human astrocyte spheroid in a high viscous methylcellulose medium and its functional advantages |
| topic | astrocytes cell immortalization differentiation status spheroid formation method |
| url | https://www.mdpi.com/2306-5354/10/3/349 |
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