Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

Abstract Multicellular tumor spheroids (MCTSs) can serve as in vitro models for solid tumors and have become widely used in basic cancer research and drug screening applications. The major challenges when studying MCTSs by optical microscopy are imaging and analysis due to light scattering within th...

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Autores principales:	Karl Olofsson, Valentina Carannante, Madoka Takai, Björn Önfelt, Martin Wiklund
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/924a88b32bdd4f598ab4d7ef11311fed
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spelling	oai:doaj.org-article:924a88b32bdd4f598ab4d7ef11311fed2021-12-02T18:53:14ZSingle cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids10.1038/s41598-021-96288-62045-2322https://doaj.org/article/924a88b32bdd4f598ab4d7ef11311fed2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96288-6https://doaj.org/toc/2045-2322Abstract Multicellular tumor spheroids (MCTSs) can serve as in vitro models for solid tumors and have become widely used in basic cancer research and drug screening applications. The major challenges when studying MCTSs by optical microscopy are imaging and analysis due to light scattering within the 3-dimensional structure. Herein, we used an ultrasound-based MCTS culture platform, where A498 renal carcinoma MCTSs were cultured, DAPI stained, optically cleared and imaged, to connect nuclear segmentation to biological information at the single cell level. We show that DNA-content analysis can be used to classify the cell cycle state as a function of position within the MCTSs. We also used nuclear volumetric characterization to show that cells were more densely organized and perpendicularly aligned to the MCTS radius in MCTSs cultured for 96 h compared to 24 h. The method presented herein can in principle be used with any stochiometric DNA staining protocol and nuclear segmentation strategy. Since it is based on a single counter stain a large part of the fluorescence spectrum is free for other probes, allowing measurements that correlate cell cycle state and nuclear organization with e.g., protein expression or drug distribution within MCTSs.Karl OlofssonValentina CarannanteMadoka TakaiBjörn ÖnfeltMartin WiklundNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Karl Olofsson Valentina Carannante Madoka Takai Björn Önfelt Martin Wiklund Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids
description	Abstract Multicellular tumor spheroids (MCTSs) can serve as in vitro models for solid tumors and have become widely used in basic cancer research and drug screening applications. The major challenges when studying MCTSs by optical microscopy are imaging and analysis due to light scattering within the 3-dimensional structure. Herein, we used an ultrasound-based MCTS culture platform, where A498 renal carcinoma MCTSs were cultured, DAPI stained, optically cleared and imaged, to connect nuclear segmentation to biological information at the single cell level. We show that DNA-content analysis can be used to classify the cell cycle state as a function of position within the MCTSs. We also used nuclear volumetric characterization to show that cells were more densely organized and perpendicularly aligned to the MCTS radius in MCTSs cultured for 96 h compared to 24 h. The method presented herein can in principle be used with any stochiometric DNA staining protocol and nuclear segmentation strategy. Since it is based on a single counter stain a large part of the fluorescence spectrum is free for other probes, allowing measurements that correlate cell cycle state and nuclear organization with e.g., protein expression or drug distribution within MCTSs.
format	article
author	Karl Olofsson Valentina Carannante Madoka Takai Björn Önfelt Martin Wiklund
author_facet	Karl Olofsson Valentina Carannante Madoka Takai Björn Önfelt Martin Wiklund
author_sort	Karl Olofsson
title	Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids
title_short	Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids
title_full	Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids
title_fullStr	Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids
title_full_unstemmed	Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids
title_sort	single cell organization and cell cycle characterization of dna stained multicellular tumor spheroids
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/924a88b32bdd4f598ab4d7ef11311fed
work_keys_str_mv	AT karlolofsson singlecellorganizationandcellcyclecharacterizationofdnastainedmulticellulartumorspheroids AT valentinacarannante singlecellorganizationandcellcyclecharacterizationofdnastainedmulticellulartumorspheroids AT madokatakai singlecellorganizationandcellcyclecharacterizationofdnastainedmulticellulartumorspheroids AT bjornonfelt singlecellorganizationandcellcyclecharacterizationofdnastainedmulticellulartumorspheroids AT martinwiklund singlecellorganizationandcellcyclecharacterizationofdnastainedmulticellulartumorspheroids
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Single cell organization and cell cycle characterization of DNA stained multicellular tumor spheroids

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