Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules
Advances in biomaterials, particularly in combination with encapsulation strategies, have provided excellent opportunities to increase reproducibility and standardization for cell culture applications. Herein, hybrid microcapsules are produced in a flow-focusing microfluidic droplet generator combin...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/11/22/3657 |
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| author | Gilles S. van Tienderen Jorke Willemse Bas van Loo Eline V. A. van Hengel Jeroen de Jonge Luc J. W. van der Laan Jeroen Leijten Monique M. A. Verstegen |
| author_facet | Gilles S. van Tienderen Jorke Willemse Bas van Loo Eline V. A. van Hengel Jeroen de Jonge Luc J. W. van der Laan Jeroen Leijten Monique M. A. Verstegen |
| author_sort | Gilles S. van Tienderen |
| collection | DOAJ |
| description | Advances in biomaterials, particularly in combination with encapsulation strategies, have provided excellent opportunities to increase reproducibility and standardization for cell culture applications. Herein, hybrid microcapsules are produced in a flow-focusing microfluidic droplet generator combined with enzymatic outside-in crosslinking of dextran-tyramine, enriched with human liver extracellular matrix (ECM). The microcapsules provide a physiologically relevant microenvironment for the culture of intrahepatic cholangiocyte organoids (ICO) and patient-derived cholangiocarcinoma organoids (CCAO). Micro-encapsulation allowed for the scalable and size-standardized production of organoids with sustained proliferation for at least 21 days in vitro. Healthy ICO (<i>n</i> = 5) expressed cholangiocyte markers, including KRT7 and KRT19, similar to standard basement membrane extract cultures. The CCAO microcapsules (<i>n</i> = 3) showed retention of stem cell phenotype and expressed LGR5 and PROM1. Furthermore, ITGB1 was upregulated, indicative of increased cell adhesion to ECM in microcapsules. Encapsulated CCAO were amendable to drug screening assays, showing a dose-response response to the clinically relevant anti-cancer drugs gemcitabine and cisplatin. High-throughput drug testing identified both pan-effective drugs as well as patient-specific resistance patterns. The results described herein show the feasibility of this one-step encapsulation approach to create size-standardized organoids for scalable production. The liver extracellular matrix-containing microcapsules can provide a powerful platform to build mini healthy and tumor tissues for potential future transplantation or personalized medicine applications. |
| first_indexed | 2024-03-09T18:25:21Z |
| format | Article |
| id | doaj.art-5faab3788a1b42398cbc800d1170178b |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T18:25:21Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-5faab3788a1b42398cbc800d1170178b2023-11-24T07:58:55ZengMDPI AGCells2073-44092022-11-011122365710.3390/cells11223657Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing MicrocapsulesGilles S. van Tienderen0Jorke Willemse1Bas van Loo2Eline V. A. van Hengel3Jeroen de Jonge4Luc J. W. van der Laan5Jeroen Leijten6Monique M. A. Verstegen7Department of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The NetherlandsDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The NetherlandsLeijten Laboratory, Department of Developmental BioEngineering, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The NetherlandsDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The NetherlandsDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The NetherlandsDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The NetherlandsLeijten Laboratory, Department of Developmental BioEngineering, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The NetherlandsDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The NetherlandsAdvances in biomaterials, particularly in combination with encapsulation strategies, have provided excellent opportunities to increase reproducibility and standardization for cell culture applications. Herein, hybrid microcapsules are produced in a flow-focusing microfluidic droplet generator combined with enzymatic outside-in crosslinking of dextran-tyramine, enriched with human liver extracellular matrix (ECM). The microcapsules provide a physiologically relevant microenvironment for the culture of intrahepatic cholangiocyte organoids (ICO) and patient-derived cholangiocarcinoma organoids (CCAO). Micro-encapsulation allowed for the scalable and size-standardized production of organoids with sustained proliferation for at least 21 days in vitro. Healthy ICO (<i>n</i> = 5) expressed cholangiocyte markers, including KRT7 and KRT19, similar to standard basement membrane extract cultures. The CCAO microcapsules (<i>n</i> = 3) showed retention of stem cell phenotype and expressed LGR5 and PROM1. Furthermore, ITGB1 was upregulated, indicative of increased cell adhesion to ECM in microcapsules. Encapsulated CCAO were amendable to drug screening assays, showing a dose-response response to the clinically relevant anti-cancer drugs gemcitabine and cisplatin. High-throughput drug testing identified both pan-effective drugs as well as patient-specific resistance patterns. The results described herein show the feasibility of this one-step encapsulation approach to create size-standardized organoids for scalable production. The liver extracellular matrix-containing microcapsules can provide a powerful platform to build mini healthy and tumor tissues for potential future transplantation or personalized medicine applications.https://www.mdpi.com/2073-4409/11/22/3657organoidsmicrocapsulesmicrofluidicsdrug screeningliver tissue engineeringcholangiocarcinoma |
| spellingShingle | Gilles S. van Tienderen Jorke Willemse Bas van Loo Eline V. A. van Hengel Jeroen de Jonge Luc J. W. van der Laan Jeroen Leijten Monique M. A. Verstegen Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules Cells organoids microcapsules microfluidics drug screening liver tissue engineering cholangiocarcinoma |
| title | Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules |
| title_full | Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules |
| title_fullStr | Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules |
| title_full_unstemmed | Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules |
| title_short | Scalable Production of Size-Controlled Cholangiocyte and Cholangiocarcinoma Organoids within Liver Extracellular Matrix-Containing Microcapsules |
| title_sort | scalable production of size controlled cholangiocyte and cholangiocarcinoma organoids within liver extracellular matrix containing microcapsules |
| topic | organoids microcapsules microfluidics drug screening liver tissue engineering cholangiocarcinoma |
| url | https://www.mdpi.com/2073-4409/11/22/3657 |
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