Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment
Monitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live b...
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MDPI AG
2022-11-01
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| author | Michael Koch Sandra Nickel Ruby Lieshout Susanna M. Lissek Martina Leskova Luc J. W. van der Laan Monique M. A. Verstegen Bruno Christ Francesco Pampaloni |
| author_facet | Michael Koch Sandra Nickel Ruby Lieshout Susanna M. Lissek Martina Leskova Luc J. W. van der Laan Monique M. A. Verstegen Bruno Christ Francesco Pampaloni |
| author_sort | Michael Koch |
| collection | DOAJ |
| description | Monitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live brightfield imaging. Growth kinetics were calculated and validated by MTS assay as well as immunohistochemistry of Ki67 to determine proliferation rates. We exposed iCCA organoids (iCCAOs) and non-tumor intrahepatic cholangiocyte organoids (ICOs) to sub-therapeutic concentrations of sorafenib. Monitoring the expansion rate of iCCAOs and ICOs revealed that iCCAO growth was inhibited by sorafenib in a time- and dose-dependent fashion, while ICOs were unaffected. Quantification of the proliferation marker Ki67 confirmed inhibition of iCCAO growth by roughly 50% after 48 h of treatment with 4 µM sorafenib. We established a robust analysis pipeline combining brightfield microscopy and a straightforward image processing approach for the label-free growth monitoring of patient-derived iCCAOs. Combined with bioanalytical validation, this approach is suitable for a fast and efficient high-throughput drug screening in tumor organoids to develop patient-specific systemic treatment options. |
| first_indexed | 2024-03-09T18:25:35Z |
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| language | English |
| last_indexed | 2024-03-09T18:25:35Z |
| publishDate | 2022-11-01 |
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| spelling | doaj.art-a40efb027cfa4c9baddd774dedc0b23b2023-11-24T07:58:13ZengMDPI AGCells2073-44092022-11-011122361310.3390/cells11223613Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib TreatmentMichael Koch0Sandra Nickel1Ruby Lieshout2Susanna M. Lissek3Martina Leskova4Luc J. W. van der Laan5Monique M. A. Verstegen6Bruno Christ7Francesco Pampaloni8Physical Biology, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, 60438 Frankfurt am Main, GermanyDepartment of Visceral, Transplant, Thoracic and Vascular Surgery, University of Leipzig Medical Center, 04103 Leipzig, GermanyDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 CN Rotterdam, The NetherlandsExperimental Medicine and Therapy Research, University of Regensburg, 93053 Regensburg, GermanyPhysical Biology, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, 60438 Frankfurt am Main, GermanyDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 CN Rotterdam, The NetherlandsDepartment of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 CN Rotterdam, The NetherlandsDepartment of Visceral, Transplant, Thoracic and Vascular Surgery, University of Leipzig Medical Center, 04103 Leipzig, GermanyPhysical Biology, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe University Frankfurt, 60438 Frankfurt am Main, GermanyMonitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live brightfield imaging. Growth kinetics were calculated and validated by MTS assay as well as immunohistochemistry of Ki67 to determine proliferation rates. We exposed iCCA organoids (iCCAOs) and non-tumor intrahepatic cholangiocyte organoids (ICOs) to sub-therapeutic concentrations of sorafenib. Monitoring the expansion rate of iCCAOs and ICOs revealed that iCCAO growth was inhibited by sorafenib in a time- and dose-dependent fashion, while ICOs were unaffected. Quantification of the proliferation marker Ki67 confirmed inhibition of iCCAO growth by roughly 50% after 48 h of treatment with 4 µM sorafenib. We established a robust analysis pipeline combining brightfield microscopy and a straightforward image processing approach for the label-free growth monitoring of patient-derived iCCAOs. Combined with bioanalytical validation, this approach is suitable for a fast and efficient high-throughput drug screening in tumor organoids to develop patient-specific systemic treatment options.https://www.mdpi.com/2073-4409/11/22/3613label-free live imagingbrightfield microscopytumor organoidsprimary liver cancerintrahepatic cholangiocarcinomasorafenib |
| spellingShingle | Michael Koch Sandra Nickel Ruby Lieshout Susanna M. Lissek Martina Leskova Luc J. W. van der Laan Monique M. A. Verstegen Bruno Christ Francesco Pampaloni Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment Cells label-free live imaging brightfield microscopy tumor organoids primary liver cancer intrahepatic cholangiocarcinoma sorafenib |
| title | Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment |
| title_full | Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment |
| title_fullStr | Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment |
| title_full_unstemmed | Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment |
| title_short | Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment |
| title_sort | label free imaging analysis of patient derived cholangiocarcinoma organoids after sorafenib treatment |
| topic | label-free live imaging brightfield microscopy tumor organoids primary liver cancer intrahepatic cholangiocarcinoma sorafenib |
| url | https://www.mdpi.com/2073-4409/11/22/3613 |
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