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
Liver three-dimensional cellular models for high-throughput chemical testing.
TL;DR: A review of 3D liver models that can be used to test drugs/chemicals for their pharmacologic and toxicologic effects, as well as the advantages and limitations of using these three-dimensional liver models for high-throughput screening is presented in this article .
Journal ArticleDOI
Improving three-dimensional human pluripotent cell culture efficiency via surface molecule coating
TL;DR: It was shown that the coating eliminated spheroid agglomeration, leading to homogenous spheoid size distribution and significant improvements in cell growth rate and volumetric yield, expected to impact large-scale hPSC production significantly.
Posted ContentDOI
Reproducibility warning: The curious case of Polyethylene glycol 6000 and spheroid cell culture
Silke Krol,Simona Serratì,Chiara Martinelli,Antonio Palazzo,Rosa Maria Iacobazzi,Maria Grazia Perrone,R. Santoliquido,Quy Khac Ong,Zhi Luo,Ahmet Bekdemir,Giulia Pinto,Ornella Cavalleri,Valentino Laquintana,Nunzio Denora,Francesco Stellacci,Annalisa Cutrignelli +15 more
TL;DR: Three-dimensional tumour spheroids or organoids present an interesting test platform for nanoparticulated drug delivery or nanoparticle toxicity, and the reproducibility of spheroid formation induced by the PEG coated surface is tested.
Book ChapterDOI
The Intersection of Biomaterials, Tissue Engineering, and Immuno-oncology
TL;DR: In this article , the authors examine advances in 3D biomaterials and tissue engineering strategies used to expand our understanding of tumor-immune interactions, and how engineered platforms are being applied toward immunotherapy development.
Proceedings ArticleDOI
Effects of combined photochemical internalization and hyperthermia are sensitively dependent on radiant exposure
TL;DR: Combination therapies of photochemical internalization (PCI) and hyperthermia (HT) were investigated in an in vitro system consisting of human glioma spheroids as mentioned 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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