High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells
Three-dimensional (3D) hepatocyte models have become a research hotspot for evaluating drug metabolism and hepatotoxicity. Compared to two-dimensional (2D) cultures, 3D cultures are better at mimicking the morphology and microenvironment of hepatocytes in vivo. However, commonly used 3D culture tech...
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MDPI AG
2024-01-01
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| Series: | International Journal of Molecular Sciences |
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| author | Yi Liu Yang Ge Yanfan Wu Yongtong Feng Han Liu Wei Cao Jinsong Xie Jingzhong Zhang |
| author_facet | Yi Liu Yang Ge Yanfan Wu Yongtong Feng Han Liu Wei Cao Jinsong Xie Jingzhong Zhang |
| author_sort | Yi Liu |
| collection | DOAJ |
| description | Three-dimensional (3D) hepatocyte models have become a research hotspot for evaluating drug metabolism and hepatotoxicity. Compared to two-dimensional (2D) cultures, 3D cultures are better at mimicking the morphology and microenvironment of hepatocytes in vivo. However, commonly used 3D culture techniques are not suitable for high-throughput drug screening (HTS) due to their high cost, complex handling, and inability to simulate cell–extracellular matrix (ECM) interactions. This article describes a method for rapid and reproducible 3D cell cultures with ECM–cell interactions based on 3D culture instrumentation to provide more efficient HTS. We developed a microsphere preparation based on a high-voltage electrostatic (HVE) field and used sodium alginate- and collagen-based hydrogels as scaffolds for 3D cultures of HepG2 cells. The microsphere-generating device enables the rapid and reproducible preparation of bioactive hydrogel microspheres. This 3D culture system exhibited better cell viability, heterogeneity, and drug-metabolizing activity than 2D and other 3D culture models, and the long-term culture characteristics of this system make it suitable for predicting long-term liver toxicity. This system improves the overall applicability of HepG2 spheroids in safety assessment studies, and this simple and controllable high-throughput-compatible method shows potential for use in drug toxicity screening assays and mechanistic studies. |
| first_indexed | 2024-03-08T09:53:09Z |
| format | Article |
| id | doaj.art-b3469f3a0e274fb3958b28e1e8cf809f |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T09:53:09Z |
| publishDate | 2024-01-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b3469f3a0e274fb3958b28e1e8cf809f2024-01-29T13:56:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-01-01252108110.3390/ijms25021081High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 CellsYi Liu0Yang Ge1Yanfan Wu2Yongtong Feng3Han Liu4Wei Cao5Jinsong Xie6Jingzhong Zhang7School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, ChinaThe CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, ChinaSchool of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, ChinaThe CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, ChinaThe CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, ChinaThe CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, ChinaThe CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, ChinaSchool of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, ChinaThree-dimensional (3D) hepatocyte models have become a research hotspot for evaluating drug metabolism and hepatotoxicity. Compared to two-dimensional (2D) cultures, 3D cultures are better at mimicking the morphology and microenvironment of hepatocytes in vivo. However, commonly used 3D culture techniques are not suitable for high-throughput drug screening (HTS) due to their high cost, complex handling, and inability to simulate cell–extracellular matrix (ECM) interactions. This article describes a method for rapid and reproducible 3D cell cultures with ECM–cell interactions based on 3D culture instrumentation to provide more efficient HTS. We developed a microsphere preparation based on a high-voltage electrostatic (HVE) field and used sodium alginate- and collagen-based hydrogels as scaffolds for 3D cultures of HepG2 cells. The microsphere-generating device enables the rapid and reproducible preparation of bioactive hydrogel microspheres. This 3D culture system exhibited better cell viability, heterogeneity, and drug-metabolizing activity than 2D and other 3D culture models, and the long-term culture characteristics of this system make it suitable for predicting long-term liver toxicity. This system improves the overall applicability of HepG2 spheroids in safety assessment studies, and this simple and controllable high-throughput-compatible method shows potential for use in drug toxicity screening assays and mechanistic studies.https://www.mdpi.com/1422-0067/25/2/1081hepatocyte3D cultureshydrogelmicrosphereshigh-voltage electrostatic fieldRNA-seq |
| spellingShingle | Yi Liu Yang Ge Yanfan Wu Yongtong Feng Han Liu Wei Cao Jinsong Xie Jingzhong Zhang High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells International Journal of Molecular Sciences hepatocyte 3D cultures hydrogel microspheres high-voltage electrostatic field RNA-seq |
| title | High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells |
| title_full | High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells |
| title_fullStr | High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells |
| title_full_unstemmed | High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells |
| title_short | High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells |
| title_sort | high voltage electrostatic field hydrogel microsphere 3d culture system improves viability and liver like properties of hepg2 cells |
| topic | hepatocyte 3D cultures hydrogel microspheres high-voltage electrostatic field RNA-seq |
| url | https://www.mdpi.com/1422-0067/25/2/1081 |
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