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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High throughput analysis of the penetration of iron oxide/polyethylene glycol nanoparticles into multicellular breast cancer tumor spheroids
TL;DR: A review of 3D Spheroid Cultures as a Tool for Biomedical and Cancer Research: Needs, Rationale and Implications for the Future, and the Engineering Journey to More Predictive Models.
Preparation and analysis of cutaneous primary myofibroblast three-dimensional culture: an experimental system for myofibroblast deactivation studies.
TL;DR: The study showed that spheroid formation does not trigger apoptosis, necrotic cell death and cyclooxygenase-2 (COX-2) protein induction associated with inflammatory response and highlights the possible relevance of clusters as reservoir of myofibroblasts for regulating my ofibroblast and tissue turnover.
Journal ArticleDOI
Anticancer Activity of Bitter Melon-Derived Vesicles Extract against Breast Cancer
TL;DR: In this paper , the authors investigated whether bitter melon-derived vesicles extract (BMVE) has antitumor activity against breast cancer and found that BMVE could dramatically inhibit tumor growth in vivo with negligible adverse effects.
Posted ContentDOI
Growth of tumor emboli within a vessel model reveals dependence on the magnitude of mechanical constraint.
Jonathan Kulwatno,Jamie Gearhart,Xiangyu Gong,Nora Herzog,Matthew Getzin,Mihaela Skobe,Kristen L. Mills +6 more
TL;DR: Light is shed on increased tumor burden in vessel-based growth and indicates a need to study tumor progression in similar environments by modelling the vessel geometry with a cylindrical microchannel in a hydrogel.
References
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Journal ArticleDOI
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.
Journal ArticleDOI
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
TL;DR: A mild method for the generation of MCTS, in which individual spheroids form in hanging drops suspended from a microtiter plate, which has applications for basic studies of physiology and metabolism, tumor biology, toxicology, cellular organization, and the development of bioartificial tissue.
Journal ArticleDOI
The use of 3-D cultures for high-throughput screening: the multicellular spheroid model.
TL;DR: 3-D in vitro systems for drug development, with a focus on screening for novel antitumor drugs, are addressed, and the advantages and limitations of these systems of intermediate complexity are discussed.
Journal ArticleDOI
Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane
TL;DR: It is reported that tissue-specific vectorial secretion coincides with the formation of functional alveoli-like structures by primary mammary epithelial cells cultured on a reconstituted basement membrane matrix (derived from Engelbreth-Holm-Swarm murine tumour), which reproduce the dual role of mammaries to secrete vectorially and to sequester milk proteins.
Journal ArticleDOI
Rotation-mediated histogenetic aggregation of dissociated cells. A quantifiable approach to cell interactions in vitro.
TL;DR: A cell aggregation procedure based on readily standardizable manipulations is described, whereby cells dissociated enzymatically from embryonic tissues may be aggregated in various combinations and concentrations into developmentally effective multicellular structures; it is suitable for analyzing mutual reactions of cells and their responses to diverse environmental conditions.
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