Rapid generation of single-tumor spheroids for high-throughput cell function and toxicity analysis.
Andrea Ivascu,Manfred Kubbies +1 more
TLDR
The authors present a rapid method to generate single spheroids in suspension culture in individual wells with homogeneous sizes, morphologies, and stratification of proliferating cells in the rim and dying Cells in the core region in a true suspension culture.Abstract:
Spheroids are widely used in biology because they provide an in vitro 3-dimensional (3D) model to study proliferation, cell death, differentiation, and metabolism of cells in tumors and the response of tumors to radiotherapy and chemotherapy. The methods of generating spheroids are limited by size heterogeneity, long cultivation time, or mechanical accessibility for higher throughput fashion. The authors present a rapid method to generate single spheroids in suspension culture in individual wells. A defined number of cells ranging from 1000 to 20,000 were seeded into wells of poly-HEMA-coated, 96-well, round-or conical-bottom plates in standard medium and centrifuged for 10 min at 1000 g. This procedure generates single spheroids in each well within a 24-h culture time with homogeneous sizes, morphologies, and stratification of proliferating cells in the rim and dying cells in the core region. Because a large number of tumor cell lines form only loose aggregates when cultured in 3D, the authors also performed a screen for medium additives to achieve a switch from aggregate to spheroid morphology. Small quantities of the basement membrane extract Matrigel, added to the culture medium prior to centrifugation, most effectively induced compact spheroid formation. The compact spheroid morphology is evident as early as 24 h after centrifugation in a true suspension culture. Twenty tumor cell lines of different lineages have been used to successfully generate compact, single spheroids with homogenous size in 96-well plates and are easily accessible for subsequent functional analysis.read more
Citations
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Development of Multicellular Tumor Spheroid (MCTS) Culture from Breast Cancer Cell and a High Throughput Screening Method Using the MTT Assay
TL;DR: A simple way of cultivating homogenous MCTS cultures with compact and rigid structure from the MCF-7 cells is reported and the MTT assay emerged as a better indicator to apoptosis event in comparison to the LDH release assay.
Journal ArticleDOI
Design of spherically structured 3D in vitro tumor models -Advances and prospects
TL;DR: This review provides an in-depth focus on current developments regarding spherically structured scaffolds engineered into in vitro 3D tumor models, and discusses future advances toward all-encompassing platforms that may provide an improved in vitro/in vivo correlation in a foreseeable future.
Journal ArticleDOI
Mammalian Target of Rapamycin Contributes to the Acquired Apoptotic Resistance of Human Mesothelioma Multicellular Spheroids
Dario Barbone,Dario Barbone,Tsung-Ming Yang,Jeffrey R. Morgan,Giovanni Gaudino,V. Courtney Broaddus +5 more
TL;DR: The three-dimensional model may reflect a more clinically relevant in vitro setting in which mTOR exhibits anti-apoptotic properties and S6K contribute to the apoptotic resistance of mesothelioma cells in three- dimensional, not in two-dimensional, cultures.
Journal ArticleDOI
High-resolution deep imaging of live cellular spheroids with light-sheet-based fluorescence microscopy
TL;DR: This review illustrates the benefits of cellular spheroids in the life sciences and suggests that LSFM is essential for investigations of cellular and sub-cellular dynamic processes in three-dimensions over time and space.
Journal ArticleDOI
Development of an Innovative 3D Cell Culture System to Study Tumour - Stroma Interactions in Non-Small Cell Lung Cancer Cells
Arno Amann,Marit Zwierzina,Gabriele Gamerith,Mario Bitsche,Julia M. Huber,Georg F. Vogel,Michael J.F. Blumer,Stefan Koeck,Elisabeth J. Pechriggl,Jens M. Kelm,Wolfgang Hilbe,Heinz Zwierzina +11 more
TL;DR: A novel 3D co-culture model using non-small cell lung cancer (NSCLC) cell lines in combination with lung fibroblasts that allows the investigation of tumour-stroma interactions and a better reflection of in vivo conditions of cancer cells in their microenvironment is described.
References
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Cell and environment interactions in tumor microregions: the multicell spheroid model
TL;DR: The special cellular microecology of tumors influences responsiveness to therapeutic agents and has implications for future directions in cancer research.
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Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types.
Jens M. Kelm,Jens M. Kelm,Nicholas E. Timmins,Catherine Brown,Martin Fussenegger,Lars K. Nielsen +5 more
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Journal ArticleDOI
Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane
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