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Single Cell Mass Cytometry of Non-Small Cell Lung Cancer Cells Reveals Complexity of In Vivo and Three-Dimensional Models over the Petri-Dish

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Avicor Ltd., H6726 Szeged, Hungary
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University of Szeged, PhD School in Biology, H6726 Szeged, Hungary
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AstridBio Technologies Ltd., H6726 Szeged, Hungary
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Laboratory of Functional Genomics, HAS BRC, H6726 Szeged, Hungary
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Avidin Ltd., H6726 Szeged, Hungary
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Research Group for Cortical Microcircuits of the Hungarian Academy of Sciences, Department of Physiology, Anatomy and Neuroscience, University of Szeged, H6726 Szeged, Hungary
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Department of Physiology, Anatomy and Neuroscience, Faculty of Science and Informatics, University of Szeged, H6726 Szeged, Hungary
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Authors to whom correspondence should be addressed.
Cells 2019, 8(9), 1093; https://doi.org/10.3390/cells8091093
Received: 31 July 2019 / Revised: 12 September 2019 / Accepted: 15 September 2019 / Published: 16 September 2019
(This article belongs to the Special Issue Single Cell Analysis)
Single cell genomics and proteomics with the combination of innovative three-dimensional (3D) cell culture techniques can open new avenues toward the understanding of intra-tumor heterogeneity. Here, we characterize lung cancer markers using single cell mass cytometry to compare different in vitro cell culturing methods: two-dimensional (2D), carrier-free, or bead-based 3D culturing with in vivo xenografts. Proliferation, viability, and cell cycle phase distribution has been investigated. Gene expression analysis enabled the selection of markers that were overexpressed: TMEM45A, SLC16A3, CD66, SLC2A1, CA9, CD24, or repressed: EGFR either in vivo or in long-term 3D cultures. Additionally, TRA-1-60, pan-keratins, CD326, Galectin-3, and CD274, markers with known clinical significance have been investigated at single cell resolution. The described twelve markers convincingly highlighted a unique pattern reflecting intra-tumor heterogeneity of 3D samples and in vivo A549 lung cancer cells. In 3D systems CA9, CD24, and EGFR showed higher expression than in vivo. Multidimensional single cell proteome profiling revealed that 3D cultures represent a transition from 2D to in vivo conditions by intermediate marker expression of TRA-1-60, TMEM45A, pan-keratin, CD326, MCT4, Gal-3, CD66, GLUT1, and CD274. Therefore, 3D cultures of NSCLC cells bearing more putative cancer targets should be used in drug screening as the preferred technique rather than the Petri-dish. View Full-Text
Keywords: single cell mass cytometry; single cell proteomics; non-small cell lung cancer; three-dimensional tissue culture single cell mass cytometry; single cell proteomics; non-small cell lung cancer; three-dimensional tissue culture
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Alföldi, R.; Balog, J.Á.; Faragó, N.; Halmai, M.; Kotogány, E.; Neuperger, P.; Nagy, L.I.; Fehér, L.Z.; Szebeni, G.J.; Puskás, L.G. Single Cell Mass Cytometry of Non-Small Cell Lung Cancer Cells Reveals Complexity of In Vivo and Three-Dimensional Models over the Petri-Dish. Cells 2019, 8, 1093.

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