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
Tumor vascularization is critical for oncolytic vaccinia virus treatment of peritoneal carcinomatosis.
Kathryn Ottolino-Perry,Kathryn Ottolino-Perry,Nan Tang,Renee Head,Calvin Ng,Rozanne Arulanandam,Fernando A. Angarita,Fernando A. Angarita,Sergio A. Acuna,Yonghong Chen,John C. Bell,Ralph S. DaCosta,J. Andrea McCart,J. Andrea McCart +13 more
TL;DR: It is demonstrated that tumor vasculature has a critical role in virus delivery and tumor response, which will have significant implications in the clinical setting, both in understanding timing of therapies and in designing combination treatment strategies.
Journal ArticleDOI
3D Spheroids' Sensitivity to Electric Field Pulses Depends on Their Size
TL;DR: It is found that small human HCT-116 colorectal spheroids are more sensitive to electric field pulses than larger ones, and characteristics of 3D cell models have to be taken into account to avoid biased conclusions of experimental data.
Book ChapterDOI
From Single Cells to Engineered and Explanted Tissues: New Perspectives in Bacterial Infection Biology.
Simone Bergmann,Michael Steinert +1 more
TL;DR: The highest level of cell culture complexity is reached by engineered and explanted tissues which currently pave the way for a more comprehensive view on microbial pathogenicity mechanisms.
Journal ArticleDOI
Increased sensitivity of glioma cells to 5-fluorocytosine following photo-chemical internalization enhanced nonviral transfection of the cytosine deaminase suicide gene.
Frederick Wang,Genesis Zamora,Chung Ho Sun,Anthony Trinidad,Changho Chun,Young Jik Kwon,Kristian Berg,Steen J. Madsen,Henry Hirschberg,Henry Hirschberg +9 more
TL;DR: Results clearly indicate that AlPcS2a-mediated PCI can be used to enhance transfection of a tumor suicide gene such as CD, in malignant glioma cells and cells transfected with both the CD and UPRT genes had a pronounced bystander effect.
Journal ArticleDOI
Focused ultrasound-mediated sonochemical internalization: an alternative to light-based therapies.
TL;DR: Examination of ultrasonic activation of sonosensitizers together with the anticancer agent bleomycin together with BLM significantly inhibits the ability of treated glioma cells to grow as three-dimensional tumor spheroids in vitro.
References
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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.
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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
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Journal ArticleDOI
Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane
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Rotation-mediated histogenetic aggregation of dissociated cells. A quantifiable approach to cell interactions in vitro.
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