دورية أكاديمية
Novel application of live imaging to determine the functional cell biology of endothelial-to-mesenchymal transition (EndMT) within a liver-on-a-chip platform.
| العنوان: | Novel application of live imaging to determine the functional cell biology of endothelial-to-mesenchymal transition (EndMT) within a liver-on-a-chip platform. |
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| المؤلفون: | Whiteford J; Barts and the London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, London, UK., Arokiasamy S; Barts and the London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, London, UK., Thompson CL; Barts and the London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, London, UK., Dufton NP; Barts and the London School of Medicine and Dentistry, William Harvey Research Institute, Queen Mary University of London, London, UK. |
| المصدر: | In vitro models [In Vitro Model] 2022; Vol. 1 (6), pp. 413-421. Date of Electronic Publication: 2022 Sep 20. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer International Publishing Country of Publication: Switzerland NLM ID: 9918506185506676 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2731-3441 (Electronic) Linking ISSN: 27313433 NLM ISO Abbreviation: In Vitro Model Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: [Cham, Switzerland] : Springer International Publishing, [2022]- |
| مستخلص: | Objective: Imaging endothelial cell behaviour under physiological conditions, particularly those associated with chronic fibrotic pathologies, is an incredibly challenging endeavour. While short-term assessments (hours) can be achieved with techniques such as intravital microscopy, vascular changes often occur over days and weeks which is unfeasible with current imaging techniques. These challenges are exemplified within the liver where liver sinusoidal endothelial cells (LSECs) are known to undergo dramatic changes termed endothelial-to-mesenchymal transition (EndMT) during fibrotic liver disease. Despite the established presence of EndMT in liver disease, the inaccessibility of viable liver tissue, and simplicity of 2D culture techniques has meant, the role of EndMT during disease progression remains largely undetermined. This study describes the development of novel fluorescent EndMT reporters to identify, track, and characterise the migratory behaviour of EndMT cells. We show that liver-on-a-chip (LOAC) platforms provide a flexible, optically accessible, and physiologically relevant microenvironment to study the vascular dynamics of EndMT during liver disease. Methods: Identification, creation, and application of an EndMT-specific fluorescent reporter construct (EndMT-Rep). Transduction of EC using lentiviral packaged CNN1-eGFP construct as an inducible EndMT-Rep (CNN1-Rep) to 2D, 3D, and 4D imaging techniques for fixed and live cell imaging. Combined application of live and fixed imaging technologies to measure EndMT using CNN1-Rep on LOAC platform under physiological conditions. Demonstration of the high-resolution single-cell EndMT tracking by live cell time-lapse microscopy and with post-acquisition processing to perform a comparative study of CNN1-Rep and healthy LSECs within a NASH-like LOAC microenvironment. Conclusions: LOAC enables prolonged, multi-platform imaging of endothelial cell sub-populations such as those undergoing EndMT in 2D and 3D cultures. Our study highlights the application of EndMT reporters, such as CNN1-Rep, to provide high-resolution imaging of EndMT behaviour for the first time under physiologically relevant liver microenvironment. Overall, these methods reveal the adaptability and impact of live-cell imaging on uncovering vascular behaviours, such as EndMT, that are unattainable in viable tissue or conventional 2D in vitro experiments. Supplementary Information: The online version contains supplementary material available at 10.1007/s44164-022-00034-9. Competing Interests: Conflict of interest N.D received an Organ-on-a-Chip Network & Emulate Proof of Concept Award providing access to Emulate LOAC and culture equipment at Queen Mary Organs-on-Chips Centre. C.L.T. is the Centre Scientist of the Queen Mary + Emulate Organs-on-Chips Centre, which is part funded by Emulate Inc. Emulate Inc. was not involved in the preparation of this manuscript. The remaining authors declare that they have no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. (© The Author(s) 2022.) |
| References: | Biol Proced Online. 2016 Apr 27;18:10. (PMID: 27127420) Br J Radiol. 2016 Aug;89(1064):20150675. (PMID: 27115318) J Hepatol. 2015 Sep;63(3):733-42. (PMID: 26055800) Hepatology. 2020 Dec;72(6):2119-2133. (PMID: 32145072) Cell Signal. 2015 Aug;27(8):1589-96. (PMID: 25917318) J Biol Chem. 2019 Oct 18;294(42):15466-15479. (PMID: 31481467) Nat Commun. 2017 Oct 12;8(1):895. (PMID: 29026072) Circ Res. 2021 Apr 16;128(8):1173-1190. (PMID: 33703914) Hepatol Commun. 2020 Nov 29;5(2):217-233. (PMID: 33553970) Nature. 2019 Nov;575(7783):512-518. (PMID: 31597160) J Clin Transl Hepatol. 2018 Jun 28;6(2):198-207. (PMID: 29951365) J Vis Exp. 2009 Oct 02;(32):. (PMID: 19801965) Sci Transl Med. 2019 Nov 6;11(517):. (PMID: 31694927) Front Med (Lausanne). 2021 Oct 20;8:750044. (PMID: 34746184) Mol Cell. 2019 Aug 8;75(3):644-660.e5. (PMID: 31398325) J Hepatol. 2016 Jul;65(1):75-83. (PMID: 27067455) |
| فهرسة مساهمة: | Keywords: EndMT; Fibrosis; Organ-on-a-chip; Vascular biology |
| تواريخ الأحداث: | Date Created: 20221226 Latest Revision: 20221227 |
| رمز التحديث: | 20221227 |
| مُعرف محوري في PubMed: | PMC9767233 |
| DOI: | 10.1007/s44164-022-00034-9 |
| PMID: | 36570669 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2731-3441 |
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| DOI: | 10.1007/s44164-022-00034-9 |