Assessment of NK cytotoxicity and interactions with porcine endothelial cells by live-cell imaging in 2D static and 3D microfluidic systems

Article Article scientifique Article Assessment of NK cytotoxicity and interactions with porcine endothelial cells by live-cell imaging in 2D static and 3D microfluidic systems eng 2024-10-15 2024-10-15 author Tran Thao Anh Department of Medicine, Laboratory of Translational Immunology, Division of Immunology and Allergy, Faculty of Medicine, Geneva University Hospitals, Geneva, Switzerland. 975133 Département de médecine (DEMED) author Galdina Viktoriia Department of Medicine, Laboratory of Translational Immunology, Division of Immunology and Allergy, Faculty of Medicine, Geneva University Hospitals, Geneva, Switzerland. 975751 Département de médecine (DEMED) author Urquidi Gandarillas Oscar 0000-0002-7406-1901 Department of Physical Chemistry, Faculty of Science, University of Geneva, Geneva, Switzerland. 931737 author Reis Galvao Daniela Department of Medicine, Laboratory of Translational Immunology, Division of Immunology and Allergy, Faculty of Medicine, Geneva University Hospitals, Geneva, Switzerland. 971814 Département de médecine (DEMED) author Rieben Robert Department for BioMedical Research (DBMR), University of Bern, Bern, Switzerland. author Adachi Takuji 0000-0002-0577-1926 Department of Physical Chemistry, Faculty of Science, University of Geneva, Geneva, Switzerland. 930973 Département de chimie physique (CHIFI) author Puga Yung Gisella 0000-0002-2283-7798 Department of Medicine, Laboratory of Translational Immunology, Division of Immunology and Allergy, Faculty of Medicine, Geneva University Hospitals, Geneva, Switzerland. gisella.pugayung@unige.ch. 666337 author Seebach Jorg Dieter Department of Medicine, Laboratory of Translational Immunology, Division of Immunology and Allergy, Faculty of Medicine, Geneva University Hospitals, Geneva, Switzerland. joerg.seebach@hug.ch. 647894 Département de médecine (DEMED) 24199 10.1038/s41598-024-75217-3 39406778 2045-2322 PMC11480498 https://www.nature.com/articles/s41598-024-75217-3 Natural Killer (NK) cells are pivotal in immune responses to viral infections, malignancies, autoimmune diseases, and transplantation. Assessment of NK cell adhesion, migration, and cytotoxicity is fundamental for in vitro studies. We propose a novel live-cell tracking method that addresses these three major aspects of NK cell function using human NK cells and primary porcine aortic endothelial cells (PAECs) in two-dimensional (2D) static assays and an in-house cylindrical 3D microfluidic system. The results showed a significant increase of NK cytotoxicity against pTNF-activated PAECs, with apoptotic cell death observed in the majority of dead cells, while no difference was observed in the conventional Delfia assay. Computed analysis of NK cell trajectories revealed distinct migratory behaviors, including trajectory length, diameter, average speed, and arrest coefficient. In 3D microfluidic experiments, NK cell attachment to pTNF-activated PAECs substantially increased, accompanied by more dead PAECs compared to control conditions. NK cell trajectories showed versatile migration in various directions and interactions with PAECs. This study uniquely demonstrates NK attachment and killing in a 3D system that mimics blood vessel conditions. Our microscope method offers sensitive single-cell level results, addressing diverse aspects of NK functions. It is adaptable for studying other immune and target cells, providing insights into various biological questions. 3D microfluidics Cell trajectory Cell-cell interaction Cytotoxicity Live cell imaging NK cell Dewey 540 Dewey 616 Swiss National Science Foundation Xeno2Cure - advanced engineering and testing of organ donor pigs 198577 CH Programmes 38 Département de chimie physique 142 56 Département de médecine 221 false 1393 Groupe Adachi 518 Cellules NK et récepteurs FcyR dans l'auto-immunité et le rejet de xénotransplantation 856 Scientific reports 14 1 1a666643-03c0-4ea5-b3e4-2c8629ca99d5 Article (Published version) Public Article_scientifique_from_crossref.pdf 4517722 application/pdf CC BY-NC-ND-4.0 018a80e325eff5da8057fe7eae27e746e44a5e4c1dd7f2c8c5e3ffc6543c37a5 aa90e7ca-273f-4095-b937-81775aacc37b Supplemental data Public 41598_2024_75217_MOESM6_ESM.docx 1668932 application/vnd.openxmlformats-officedocument.wordprocessingml.document 1a892de827afdc1a04a7f49a558e0ca7b54fe9e17707814ee8e1f8aa23c6a12b 595d0bb3-d60e-472f-a1e6-8a58103090a1 Supplemental data Public 41598_2024_75217_MOESM8_ESM.docx 33056 application/vnd.openxmlformats-officedocument.wordprocessingml.document d0df31d40b99be2e5b401d37eb9f68c76e0513b02468664594bde992aa741558 openAccess Faculté des sciences / Section de chimie et biochimie / Département de chimie physique Faculté de médecine / Section de médecine clinique / Département de médecine

Title	Published in	Year	Views	Downloads

Cytotoxic Responses Mediated by NK Cells and Cytotoxic T Lymphocytes in Xenotransplantation	Transplant international	2025	82	62
Assessment of NK cytotoxicity and interactions with porcine endothelial cells by live-cell imaging in 2D static and 3D microfluidic systems	Scientific reports	2024	93	44

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