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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Mechanical properties of cell sheets and spheroids: the link between single cells and complex tissues.
Yuri M. Efremov,I.M. Zurina,Viktoria S. Presniakova,Nastasia V. Kosheleva,Denis Butnaru,Andrey A. Svistunov,Yury Rochev,Yury Rochev,Peter S. Timashev +8 more
TL;DR: In this paper, the authors review the data on mechanical characterization of cell sheets and spheroids, focusing both on advances in the measurement techniques and current understanding of the subject, and suggest that interplay between the ECM, intercellular junctions, and cellular contractility determines the behavior and mechanical properties of the cell aggregates.
Journal ArticleDOI
Let-7 microRNA controls invasion-promoting lysosomal changes via the oncogenic transcription factor myeloid zinc finger-1.
Siri Amanda Tvingsholm,Malene Hansen,Knut Kristoffer Bundgaard Clemmensen,Ditte Marie Brix,Bo Rafn,Lisa B. Frankel,Riku Louhimo,José M.A. Moreira,Sampsa Hautaniemi,Irina Gromova,Marja Jäättelä,Tuula Kallunki +11 more
TL;DR: Let-7 microRNA, a well-known tumor suppressor in breast cancer, is identified as a direct negative regulator of Myeloid Zinc Finger1, and it is shown that MZF1 regulates lysosome trafficking in ErbB2-positive breast cancer cells.
Journal ArticleDOI
Photochemical internalization-mediated nonviral gene transfection: Polyamine core-shell nanoparticles as gene carrier
Genesis Zamora,Frederick Wang,Chung-Ho Sun,Anthony Trinidad,Young Jik Kwon,Soo Kyung Cho,Kristian Berg,Steen J. Madsen,Henry Hirschberg +8 more
TL;DR: The results clearly demonstrated that the PCI can enhance the delivery of both the PTEN or CD genes in human glioma cell monolayers and multicell tumor spheroids.
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Optimization of invasion-specific effects of betulin derivatives on prostate cancer cells through lead development
Ville Härmä,Raisa Haavikko,Johannes Virtanen,Ilmari Ahonen,Hannu-Pekka Schukov,Sami Alakurtti,Enkhee Purev,Heiko Rischer,Jari Yli-Kauhaluoma,Vânia M. Moreira,Matthias Nees,Kirsi-Marja Oksman-Caldentey +11 more
TL;DR: Target de-convolution studies, combined with the phenotypic analyses of multicellular organoids in 3D models, revealed specific inhibition of AKT signaling linked to effects on the organization of the actin cytoskeleton as the most likely driver of altered cell morphology and motility.
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Perspective: Flicking with flow: Can microfluidics revolutionize the cancer research?
Tamal Das,Suman Chakraborty +1 more
TL;DR: The biophysical aspects of cancer are described and how microfluidic system could be useful in developing a deeper understanding on the underlying mechanisms, and the effects of the confined environment of microchannel on cellular dynamics are illustrated.
References
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