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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Journal ArticleDOI
Developing membrane-derived nanocarriers for ex vivo therapy of homologous breast cancer cells.
TL;DR: In this article, the authors developed biomimetic nanocarriers for specific homologous targeting of the anticancer drugs ammonium pyrrolidine dithiocarbamate (PDTC) and doxorubicin.
Book ChapterDOI
Human Microtumors Generated in 3D: Novel Tools for Integrated In Situ Studies of Cancer Immunotherapies.
Lothar Hambach,Lothar Hambach,Andreas Buser,Marcel Vermeij,Nadine Pouw,Theo H. van der Kwast,Els Goulmy +6 more
TL;DR: A novel assay for multiparametric in situ analysis of therapeutic effects on individual human three-dimensional tumors, allowing integrated analyses of the antitumor efficacy of cancer specific immunotherapy in situ.
Dissertation
Impedance-based analysis of 3D tissue models: A novel measurement setup for novel measurement modes
TL;DR: The characterization and testing of a new impedance-based flow channel is a label-free, non-invasive and time resolved new readout technique to monitor 3D tissue models and their changes within biomedical experiments.
Dissertation
Effects on migration of breast cancer cells in 3D models upon MMP inhibition
TL;DR: In this paper, the authors proposed a method to solve the problem of the problem: this paper..s.p.a.d.t.c.t..
Journal ArticleDOI
Three-dimensional cell cultures as preclinical models to assess the biological activity of phytochemicals in breast Cancer.
Nádia Calvo Martins Okuyama,Diego Luis Ribeiro,Cláudia Quintino da Rocha,Érica Romão Pereira,Ilce Mara de Syllos Cólus,Juliana Mara Serpeloni +5 more
TL;DR: A review of the most common 3D cell culture methods, focusing on multicellular tumor spheroids (MCTS) of breast cancer cell lines used in the discovery of phytochemicals with anticancer properties in the last ten years is presented in this paper .
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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