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
More filters
Journal ArticleDOI
Macrophages as Cell-Based Delivery Systems for Nanoshells in Photothermal Therapy
TL;DR: Of particular interest is the observation that, after near-infrared exposure of spheroids containing nanoshell-loaded macrophages, sufficient heat was generated to suppress spheroid growth.
Journal ArticleDOI
High-Content Monitoring of Drug Effects in a 3D Spheroid Model.
Frédérique Mittler,Patricia Obeid,Anastasiia V. Rulina,Anastasiia V. Rulina,Vincent Haguet,Xavier Gidrol,Maxim Y Balakirev +6 more
TL;DR: It is demonstrated that the use of a spheroid model may be straightforward and can provide information that is not directly available with a standard 2D approach, and once fully validated in a wider range of conditions, this method could be particularly valuable for phenotype-based drug discovery.
Journal ArticleDOI
Generation of Homogenous Three-Dimensional Pancreatic Cancer Cell Spheroids Using an Improved Hanging Drop Technique.
Matthew Ware,Matthew Ware,Kevin L. Colbert,Vazrik Keshishian,Jason C. Ho,Stuart J. Corr,Steven A. Curley,Biana Godin +7 more
TL;DR: The aim of this study was to design and test an easy-to-use and highly reproducible modification of the hanging drop method for tumor sphere formation by adding methylcellulose polymer and believe that this method can be used as a reliable and reproducible technique to make 3D cancer spheroids for use in tumor biology research and evaluation of therapeutic responses, and for the development of bio-artificial tissues.
Journal ArticleDOI
Incorporation of multicellular spheroids into 3-D polymeric scaffolds provides an improved tumor model for screening anticancer drugs
Won Jin Ho,Edward A. Pham,Jun W. Kim,Christopher W. Ng,Jae H. Kim,Daniel T. Kamei,Benjamin M. Wu +6 more
TL;DR: A novel in vitro model in which multicellular aggregates, or spheroids, were incorporated into 3‐D porous scaffolds showed that spheroid‐seeded scaffolds have much higher drug resistance than monolayer cultures, spheroid cultures on flat substrates,or scaffolds seeded with dispersed cells.
Journal ArticleDOI
Effect of Shape, Size, and Aspect Ratio on Nanoparticle Penetration and Distribution inside Solid Tissues Using 3D Spheroid Models
Rachit Agarwal,Patrick Jurney,Mansi Raythatha,Vikramjit Singh,Sidlgata V. Sreenivasan,Li Shi,Krishnendu Roy +6 more
TL;DR: Experimental and theoretical studies are reported on how nanoscale geometry of Jet and Flash Imprint Lithography‐fabricated, polyethylene‐glycol‐based anionic nanohydrogels affect their penetration and distribution inside 3D spheroids, a model representing the intervascular region of solid, tumor‐like tissues.
References
More filters
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.
Related Papers (5)
Recent advances in three-dimensional multicellular spheroid culture for biomedical research
Ruei-Zhen Lin,Hwan-You Chang +1 more