Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs
Abstract Objectives Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unclear. This study thus aimed...
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| Format: | Article |
| Language: | English |
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BMC
2022-05-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-02850-y |
| _version_ | 1818191484142223360 |
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| author | Mitsuru Mizuno Takahisa Matsuzaki Nobutake Ozeki Hisako Katano Hideyuki Koga Takanori Takebe Hiroshi Y. Yoshikawa Ichiro Sekiya |
| author_facet | Mitsuru Mizuno Takahisa Matsuzaki Nobutake Ozeki Hisako Katano Hideyuki Koga Takanori Takebe Hiroshi Y. Yoshikawa Ichiro Sekiya |
| author_sort | Mitsuru Mizuno |
| collection | DOAJ |
| description | Abstract Objectives Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unclear. This study thus aimed to identify the biological and physical factors involved in the differences in freeze–thaw tolerance between MSCs and HUVECs. Materials and methods For biological analysis, MSC and HUVEC viability after freeze-thawing and alteration of gene expression in response to dimethyl sulfoxide (DMSO, a cryoprotectant) were quantitatively evaluated. For physical analysis, the cell membrane fluidity of MSCs and HUVECs before and after DMSO addition was assessed using a histogram for generalized polarization frequency. Results HUVECs showed lower live cell rates and higher gene expression alteration related to extracellular vesicles in response to DMSO than MSCs. Fluidity measurements revealed that the HUVEC membrane was highly fluidic and sensitive to DMSO compared to that of MSCs. Addition of CAY10566, an inhibitor of stearoyl-coA desaturase (SCD1) that produces highly fluidic desaturated fatty acids, decreased the fluidity of HUVECs and increased their tolerance to DMSO. The combination of CAY10566 and antioxidant glutathione (GSH) treatment improved HUVEC viability from 57 to 69%. Membrane fluidity alteration may thus contribute to pore-induced DMSO influx into the cytoplasm and reactive oxygen species production, leading to greater cytotoxicity in HUVECs, which have low antioxidant capacity. Conclusions Differences in freeze–thaw tolerance originate from differences in the cell membranes with respect to fluidity and antioxidant capacity. These findings provide a basis for analyzing cell biology and membrane-physics to establish appropriate long-term preservation methods aimed at promoting transplantation therapies. |
| first_indexed | 2024-12-12T00:15:20Z |
| format | Article |
| id | doaj.art-e95b65d2ba6046aeab8ac7674489d4b2 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-12T00:15:20Z |
| publishDate | 2022-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-e95b65d2ba6046aeab8ac7674489d4b22022-12-22T00:44:51ZengBMCStem Cell Research & Therapy1757-65122022-05-0113111610.1186/s13287-022-02850-yCell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECsMitsuru Mizuno0Takahisa Matsuzaki1Nobutake Ozeki2Hisako Katano3Hideyuki Koga4Takanori Takebe5Hiroshi Y. Yoshikawa6Ichiro Sekiya7Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University (TMDU)Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama UniversityCenter for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University (TMDU)Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University (TMDU)Department of Joint Surgery and Sports Medicine, Graduate School, Tokyo Medical and Dental University (TMDU)Institute of Research, Tokyo Medical and Dental University (TMDU)Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama UniversityCenter for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University (TMDU)Abstract Objectives Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unclear. This study thus aimed to identify the biological and physical factors involved in the differences in freeze–thaw tolerance between MSCs and HUVECs. Materials and methods For biological analysis, MSC and HUVEC viability after freeze-thawing and alteration of gene expression in response to dimethyl sulfoxide (DMSO, a cryoprotectant) were quantitatively evaluated. For physical analysis, the cell membrane fluidity of MSCs and HUVECs before and after DMSO addition was assessed using a histogram for generalized polarization frequency. Results HUVECs showed lower live cell rates and higher gene expression alteration related to extracellular vesicles in response to DMSO than MSCs. Fluidity measurements revealed that the HUVEC membrane was highly fluidic and sensitive to DMSO compared to that of MSCs. Addition of CAY10566, an inhibitor of stearoyl-coA desaturase (SCD1) that produces highly fluidic desaturated fatty acids, decreased the fluidity of HUVECs and increased their tolerance to DMSO. The combination of CAY10566 and antioxidant glutathione (GSH) treatment improved HUVEC viability from 57 to 69%. Membrane fluidity alteration may thus contribute to pore-induced DMSO influx into the cytoplasm and reactive oxygen species production, leading to greater cytotoxicity in HUVECs, which have low antioxidant capacity. Conclusions Differences in freeze–thaw tolerance originate from differences in the cell membranes with respect to fluidity and antioxidant capacity. These findings provide a basis for analyzing cell biology and membrane-physics to establish appropriate long-term preservation methods aimed at promoting transplantation therapies.https://doi.org/10.1186/s13287-022-02850-yCell membrane fluidityROS resistanceCryopreserveMesenchymal stem cellsHuman umbilical vein endothelial cells |
| spellingShingle | Mitsuru Mizuno Takahisa Matsuzaki Nobutake Ozeki Hisako Katano Hideyuki Koga Takanori Takebe Hiroshi Y. Yoshikawa Ichiro Sekiya Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs Stem Cell Research & Therapy Cell membrane fluidity ROS resistance Cryopreserve Mesenchymal stem cells Human umbilical vein endothelial cells |
| title | Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs |
| title_full | Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs |
| title_fullStr | Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs |
| title_full_unstemmed | Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs |
| title_short | Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs |
| title_sort | cell membrane fluidity and ros resistance define dmso tolerance of cryopreserved synovial mscs and huvecs |
| topic | Cell membrane fluidity ROS resistance Cryopreserve Mesenchymal stem cells Human umbilical vein endothelial cells |
| url | https://doi.org/10.1186/s13287-022-02850-y |
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