Development of Multicellular Tumor Spheroid (MCTS) Culture from Breast Cancer Cell and a High Throughput Screening Method Using the MTT Assay
Open accessAbstract
In comparison to monolayer cells, MCTS has been claimed as more suitable candidate for studying drug penetration due to the high resemblance to solid tumors. However, the cultivation of MCTS is cumbersome, time consuming, and most technique fail to generate spheroids with uniform sizes. Therefore, the application of spheroid cultures in high throughput screening has been rather limiting. Besides, the lack of a well established screening protocol method that is applicable to spheroid could also be attributed to this limitation. Here we report a simple way of cultivating homogenous MCTS cultures with compact and rigid structure from the MCF-7 cells. Besides, we had also made some modifications to the standard MTT assay to realize high throughput screening of these spheroids. Using the modified protocol, tamoxifen showed cytotoxicity effect towards MCTS cultures from MCF-7 with high consistency. The results correlated well with the cultures' response assessed by LDH release assay but the latter assay was not ideal for detecting a wide range of cytotoxicity due to high basal background reading. The MTT assay emerged as a better indicator to apoptosis event in comparison to the LDH release assay. Therefore, the method for spheroid generation and the modified MTT assay we reported here could be potentially applied to high throughput screening for response of spheroid cultures generated from MCF-7 as well as other cancer cell lines towards cytotoxic stimuli.
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There is an association between size and the formation of oxygen limited environments (hypoxia) within the spheroid system.
“Our study highlights the important association between size and the formation of oxygen limited environments (hypoxia) within the spheroid system, a finding which has already been illustrated in the literature for tumour derived spheroid models [49, 50] but nevertheless has remained unknown till now in the non tumour models.”
From Direct Measurements of Oxygen Gradients in Spheroid Culture System Using Electron Parametric Resonance Oximetry (2016) · cites this paper as [49]
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References 29 · Cited by 173
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- Authors
- Ho, W. Y., Yeap, S. K., Ho, C. L., Rahim, R. A., & Alitheen, N. B.
- Affiliations
- Universiti Putra Malaysia (MY)
- Corresponding author
- Alitheen, Noorjahan Banu
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2012-09-06
- Volume
- 7
- Issue
- 9
- Pages
- e44640
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC3435305
- OpenAlex
- W2015963176
- Cited by
- 173
- References
- 29
- Citations by year
- 2026: 7 · 2025: 7 · 2024: 10 · 2023: 9 · 2022: 7 · 2021: 18 · 2020: 19 · 2019: 14 · 2018: 14 · 2017: 18 · 2016: 18 · 2015: 18 · 2014: 10 · 2013: 4
- FWCI
- 5.07
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Greater South Information System
- Funders
- Universiti Putra Malaysia
- Malaysia Toray Science Foundation
- Toray Science Foundation
- Topics
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Physical Sciences › Mathematics › Modeling and Simulation › Mathematical Biology Tumor Growth
- Keywords
- tumor spheroids, breast cancer, MCF-7 cells, high-throughput screening, drug penetration, spheroid culture, apoptosis detection
- MeSH terms
- Breast Neoplasms, Female, Humans, L-Lactate Dehydrogenase, Microscopy, Electron, Scanning, Tamoxifen, Cell Death, Antineoplastic Agents, Hormonal, High-Throughput Screening Assays, MCF-7 Cells
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