Deterministic culturing of single cells in 3D

Abstract Models using 3D cell culture techniques are increasingly accepted as the most biofidelic in vitro representations of tissues for research. These models are generated using biomatrices and bulk populations of cells derived from tissues or cell lines. We present an alternate method to culture...

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Autores principales:	Rohil Jain, Shirisha Chittiboyina, Chun-Li Chang, Sophie A. Lelièvre, Cagri A. Savran
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b79e8c355aa34c89ab658a1182d4d1eb
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spelling	oai:doaj.org-article:b79e8c355aa34c89ab658a1182d4d1eb2021-12-02T16:31:42ZDeterministic culturing of single cells in 3D10.1038/s41598-020-67674-32045-2322https://doaj.org/article/b79e8c355aa34c89ab658a1182d4d1eb2020-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-67674-3https://doaj.org/toc/2045-2322Abstract Models using 3D cell culture techniques are increasingly accepted as the most biofidelic in vitro representations of tissues for research. These models are generated using biomatrices and bulk populations of cells derived from tissues or cell lines. We present an alternate method to culture individually selected cells in relative isolation from the rest of the population under physiologically relevant matrix conditions. Matrix gel islands are spotted on a cell culture dish to act as support for receiving and culturing individual single cells; a glass capillary-based microfluidic setup is used to extract each desired single cell from a population and seed it on top of an island. Using examples of breast and colorectal cancers, we show that individual cells evolve into tumors or aspects of tumors displaying different characteristics of the initial cancer type and aggressiveness. By implementing a morphometry assay with luminal A breast cancer, we demonstrate the potential of the proposed approach to study phenotypic heterogeneity. Results reveal that intertumor heterogeneity increases with time in culture and that varying degrees of intratumor heterogeneity may originate from individually seeded cells. Moreover, we observe that a positive relationship exists between fast growing tumors and the size and heterogeneity of their nuclei.Rohil JainShirisha ChittiboyinaChun-Li ChangSophie A. LelièvreCagri A. SavranNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rohil Jain Shirisha Chittiboyina Chun-Li Chang Sophie A. Lelièvre Cagri A. Savran Deterministic culturing of single cells in 3D
description	Abstract Models using 3D cell culture techniques are increasingly accepted as the most biofidelic in vitro representations of tissues for research. These models are generated using biomatrices and bulk populations of cells derived from tissues or cell lines. We present an alternate method to culture individually selected cells in relative isolation from the rest of the population under physiologically relevant matrix conditions. Matrix gel islands are spotted on a cell culture dish to act as support for receiving and culturing individual single cells; a glass capillary-based microfluidic setup is used to extract each desired single cell from a population and seed it on top of an island. Using examples of breast and colorectal cancers, we show that individual cells evolve into tumors or aspects of tumors displaying different characteristics of the initial cancer type and aggressiveness. By implementing a morphometry assay with luminal A breast cancer, we demonstrate the potential of the proposed approach to study phenotypic heterogeneity. Results reveal that intertumor heterogeneity increases with time in culture and that varying degrees of intratumor heterogeneity may originate from individually seeded cells. Moreover, we observe that a positive relationship exists between fast growing tumors and the size and heterogeneity of their nuclei.
format	article
author	Rohil Jain Shirisha Chittiboyina Chun-Li Chang Sophie A. Lelièvre Cagri A. Savran
author_facet	Rohil Jain Shirisha Chittiboyina Chun-Li Chang Sophie A. Lelièvre Cagri A. Savran
author_sort	Rohil Jain
title	Deterministic culturing of single cells in 3D
title_short	Deterministic culturing of single cells in 3D
title_full	Deterministic culturing of single cells in 3D
title_fullStr	Deterministic culturing of single cells in 3D
title_full_unstemmed	Deterministic culturing of single cells in 3D
title_sort	deterministic culturing of single cells in 3d
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/b79e8c355aa34c89ab658a1182d4d1eb
work_keys_str_mv	AT rohiljain deterministicculturingofsinglecellsin3d AT shirishachittiboyina deterministicculturingofsinglecellsin3d AT chunlichang deterministicculturingofsinglecellsin3d AT sophiealelievre deterministicculturingofsinglecellsin3d AT cagriasavran deterministicculturingofsinglecellsin3d
_version_	1718383895307616256

Deterministic culturing of single cells in 3D

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