Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture
Rho–associated protein kinase (ROCK) inhibitors are used for the survival of single-dissociated human induced pluripotent stem cells (hiPSCs); however, their effects on the growth behaviors of hiPSCs in suspension culture are unexplored. Therefore, we investigated the effect of ROCK inhibitor on gro...
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| Format: | Article |
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MDPI AG
2022-10-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/9/11/613 |
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| author | Takaki Matsumoto Mee-Hae Kim Masahiro Kino-oka |
| author_facet | Takaki Matsumoto Mee-Hae Kim Masahiro Kino-oka |
| author_sort | Takaki Matsumoto |
| collection | DOAJ |
| description | Rho–associated protein kinase (ROCK) inhibitors are used for the survival of single-dissociated human induced pluripotent stem cells (hiPSCs); however, their effects on the growth behaviors of hiPSCs in suspension culture are unexplored. Therefore, we investigated the effect of ROCK inhibitor on growth behaviors of two hiPSC lines (Tic and 1383D2) with different formation of aggregate that attached between single cells in suspension culture. The apparent specific growth rate by long-term exposure to Y-27632, a ROCK inhibitor, was maintained throughout the culture. Long-term exposure to ROCK inhibitor led to an increase in cell division throughout the culture in both lines. Immunofluorescence staining confirmed that hiPSCs forming spherical aggregates showed localization of collagen type I on its periphery. In addition, phosphorylated myosin (pMLC) was localized at the periphery in culture under short-term exposure to ROCK inhibitor, whereas pMLC was not detected at whole the aggregate in culture under long-term exposure. Scanning electron microscopy indicated that long-term exposure to ROCK inhibitor blocked the structural alteration on the surface of cell aggregates. These results indicate that pMLC inhibition by long-term ROCK inhibition leads to enhanced growth abilities of hiPSCs in suspension culture by maintaining the structures of extracellular matrices. |
| first_indexed | 2024-03-09T19:16:17Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-09T19:16:17Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
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| series | Bioengineering |
| spelling | doaj.art-912ffb7474644c3f826786e0534133c02023-11-24T03:46:04ZengMDPI AGBioengineering2306-53542022-10-0191161310.3390/bioengineering9110613Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension CultureTakaki Matsumoto0Mee-Hae Kim1Masahiro Kino-oka2Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Osaka, JapanDepartment of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Osaka, JapanDepartment of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Osaka, JapanRho–associated protein kinase (ROCK) inhibitors are used for the survival of single-dissociated human induced pluripotent stem cells (hiPSCs); however, their effects on the growth behaviors of hiPSCs in suspension culture are unexplored. Therefore, we investigated the effect of ROCK inhibitor on growth behaviors of two hiPSC lines (Tic and 1383D2) with different formation of aggregate that attached between single cells in suspension culture. The apparent specific growth rate by long-term exposure to Y-27632, a ROCK inhibitor, was maintained throughout the culture. Long-term exposure to ROCK inhibitor led to an increase in cell division throughout the culture in both lines. Immunofluorescence staining confirmed that hiPSCs forming spherical aggregates showed localization of collagen type I on its periphery. In addition, phosphorylated myosin (pMLC) was localized at the periphery in culture under short-term exposure to ROCK inhibitor, whereas pMLC was not detected at whole the aggregate in culture under long-term exposure. Scanning electron microscopy indicated that long-term exposure to ROCK inhibitor blocked the structural alteration on the surface of cell aggregates. These results indicate that pMLC inhibition by long-term ROCK inhibition leads to enhanced growth abilities of hiPSCs in suspension culture by maintaining the structures of extracellular matrices.https://www.mdpi.com/2306-5354/9/11/613human induced pluripotent stem cellsuspension cultureRho–associated kinase inhibitoraggregate formationgrowth behaviorcollagen type I |
| spellingShingle | Takaki Matsumoto Mee-Hae Kim Masahiro Kino-oka Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture Bioengineering human induced pluripotent stem cell suspension culture Rho–associated kinase inhibitor aggregate formation growth behavior collagen type I |
| title | Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture |
| title_full | Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture |
| title_fullStr | Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture |
| title_full_unstemmed | Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture |
| title_short | Effect of Rho–Associated Kinase Inhibitor on Growth Behaviors of Human Induced Pluripotent Stem Cells in Suspension Culture |
| title_sort | effect of rho associated kinase inhibitor on growth behaviors of human induced pluripotent stem cells in suspension culture |
| topic | human induced pluripotent stem cell suspension culture Rho–associated kinase inhibitor aggregate formation growth behavior collagen type I |
| url | https://www.mdpi.com/2306-5354/9/11/613 |
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