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
The third dimension bridges the gap between cell culture and live tissue
TL;DR: It is believed that 3D cultures will have a strong impact on drug screening and will also decrease the use of laboratory animals, for example, in the context of toxicity assays.
Journal ArticleDOI
Multicellular tumor spheroids: an underestimated tool is catching up again
Franziska Hirschhaeuser,Heike Menne,Claudia Dittfeld,Jonathan West,Wolfgang Mueller-Klieser,Leoni A. Kunz-Schughart +5 more
TL;DR: The rationale, potential and flexibility of tumor spheroid mono- and cocultures for implementation into state of the art anti-cancer therapy test platforms are highlighted and the relevance of the cancer stem cell hypothesis for cancer cure is highlighted.
Journal ArticleDOI
Spheroid-based drug screen: considerations and practical approach
TL;DR: This work describes a standardized setup for reproducible, easy-handling culture, treatment and routine analysis of multicellular spheroids, the classical 3D culture system resembling many aspects of the pathophysiological situation in human tumor tissue and provides a list of human carcinoma cell lines that produce treatable sp Heroids under identical culture conditions.
Journal ArticleDOI
Three-dimensional cell culture: the missing link in drug discovery.
TL;DR: 2D and 3D culture approaches are reviewed and the strengths and relevance of each method are considered in the context of anti-cancer drug screening.
Journal ArticleDOI
Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy.
TL;DR: The suitability of spheroids as an in vitro platform for testing drug delivery systems is examined and the assay techniques required for the characterization of drug delivery and efficacy in sp Heroids are discussed.
References
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Journal Article
Oxygenation and differentiation in multicellular spheroids of human colon carcinoma.
TL;DR: The PO2 profiles of both of these human tumor spheroid types were often continuously curving with a very shallow gradient in the inner edge of the viable rim of cells surrounding the necrotic center, which has not previously been seen in other poorly differentiated human or rodent tumor spheroids.
Journal ArticleDOI
Tumor biology and experimental therapeutics
TL;DR: Research efforts should be enhanced mainly in the fields of tumor tissue modeling by heterotypic three-dimensional (3D) cultures and of apoptotic versus necrotic cell death and spheroids should become mandatory test systems in therapeutic screening programs.
Book ChapterDOI
Liquid-Overlay Culture of Cellular Spheroids
J. Carlsson,J. M. Yuhas +1 more
TL;DR: Large variations in growth rate, cell morphology, thickness of the viable cell layer and PO2 profiles were found among the tested spheroids, which parallels the variability seen in solid tumors.
Radiation response of multicell spheroids--an in vitro tumour model.
Sutherland Rm,Durand Re +1 more
TL;DR: Multicell spheroids provide an in vitro tumour model of intermediate complexity between tumours and standard cell cultures and methods were developed to study in situ repair and repopulation processes after ionizing radiation or other cytotoxic agents.
Journal ArticleDOI
A Multi-component Radiation Survival Curve Using an in Vitro Tumour Model
TL;DR: In this paper, a multi-component radiation survival curve using an in-vivo Tumour Model is presented, and the model is used to predict the survival rate of cancer patients.
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