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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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: In this article, the authors modeled a lymphatic vessel as a 200 μm-diameter channel in either a stiff or soft, bioinert agarose matrix to create a vessel-like constraint model (VLCM), and modeled colon or breast cancer tumor emboli with aggregates of HCT116 or SUM149PT cells.
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Morphometrical, Morphological, and Immunocytochemical Characterization of a Tool for Cytotoxicity Research: 3D Cultures of Breast Cell Lines Grown in Ultra-Low Attachment Plates
Fernanda Malhão,Ana Catarina Fernandes Pereira Rebelo de Macedo,Alice A. Ramos,Eduardo Rocha +3 more
TL;DR: The presented characterization of the MCAs in non-exposed conditions provided a good baseline to evaluate the cytotoxic effects of potential anticancer compounds and showed a random distribution of the proliferating and apoptotic cells throughout theMCAs, not fitting in the traditional spheroid model.
Journal ArticleDOI
3D Breast Tumor Models for Radiobiology Applications.
Akhilandeshwari Ravichandran,Julien Clegg,Mark N. Adams,Madison Hampson,Andrew Fielding,Laura J. Bray +5 more
TL;DR: In this paper, the effects of radiation therapy on normal and malignant breast cells and tissues, and explore the emerging opportunities that pre-clinical 3D models offer in improving our understanding of this treatment modality.
Journal ArticleDOI
Cytotoxicity of multicellular cancer spheroids, antibacterial, and antifungal of selected sulfonamide derivatives coupled with a salicylamide and/or anisamide scaffold
Alaaeldin M. F. Galal,Walid Fayad,Walaa S. A. Mettwally,Sanaa K Gomaa,Esam R. Ahmed,Heba A. El-Refai,Atef G. Hanna +6 more
TL;DR: The most active sulfonamide derivatives coupled with a salicylamide and/or anisamide scaffold were screened for cytotoxicity on two human cancer cell line spheroids and could be subjected to in vivo investigation as new drugs.
Journal ArticleDOI
Predicting drug sensitivity by 3D cell culture models
TL;DR: Three-dimensional (3D) cell culture may represents an modern alternative to bridge the gap between 2D cell cultures and animal models to predict drug efficacy in patients.
References
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