Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types
Abstract
Multicellular tumor spheroids (MCTS) are used as organotypic models of normal and solid tumor tissue. Traditional techniques for generating MCTS, such as growth on nonadherent surfaces, in suspension, or on scaffolds, have a number of drawbacks, including the need for manual selection to achieve a homogeneous population and the use of nonphysiological matrix compounds. In this study we describe a mild method for the generation of MCTS, in which individual spheroids form in hanging drops suspended from a microtiter plate. The method has been successfully applied to a broad range of cell lines and shows nearly 100% efficiency (i.e., one spheroid per drop). Using the hepatoma cell line, HepG2, the hanging drop method generated well‐rounded MCTS with a narrow size distribution (coefficient of variation [CV] 10% to 15%, compared with 40% to 60% for growth on nonadherent surfaces). Structural analysis of HepG2 and a mammary gland adenocarcinoma cell line, MCF‐7, composed spheroids, revealed highly organized, three‐dimensional, tissue‐like structures with an extensive extracellular matrix. The hanging drop method represents an attractive alternative for MCTS production, because it is mild, can be applied to a wide variety of cell lines, and can produce spheroids of a homogeneous size without the need for sieving or manual selection. The method has applications for basic studies of physiology and metabolism, tumor biology, toxicology, cellular organization, and the development of bioartificial tissue. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 173–180, 2003.
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Hanging drop methods have limited working volumes of 20-50 μL.
“Hanging drop methods have limited working volumes of 20–50 μL.”
From The Tumor-on-Chip: Recent Advances in the Development of Microfluidic Systems to Recapitulate the Physiology of Solid Tumors (2019) · cites this paper as [149]
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- Authors
- Kelm, J. M., Timmins, N. E., Brown, C. J., Fussenegger, M., & Nielsen, L. K.
- Affiliations
- The University of Queensland (AU)
- ETH Zurich (CH)
- Corresponding author
- Nielsen, Lars K.
- Venue
- Biotechnology and Bioengineering
- Publisher
- Wiley
- Published
- 2003-05-14
- Volume
- 83
- Issue
- 2
- Pages
- 173-180
- ISSN
- 0006-3592 (ISSN-L)
- 1097-0290
- OpenAlex
- W2119698183
- Cited by
- 934
- References
- 1
- Citations by year
- 2026: 21 · 2025: 28 · 2024: 31 · 2023: 51 · 2022: 55 · 2021: 60 · 2020: 72 · 2019: 58 · 2018: 70 · 2017: 67 · 2016: 72 · 2015: 52 · 2014: 38 · 2013: 48 · 2012: 38
- FWCI
- 11.30
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biotechnology and Bioengineering (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Physical Sciences › Mathematics › Modeling and Simulation › Mathematical Biology Tumor Growth
- Keywords
- tumor spheroids, hanging drop method, HepG2 cells, extracellular matrix, bioartificial tissues
- MeSH terms
- Breast Neoplasms, Female, Carcinoma, Hepatocellular, Humans, Liver Neoplasms, Microscopy, Electron, Microscopy, Electron, Scanning, Models, Biological, Neoplasms, Tumor Cells, Cultured, Cell Death, Cell Culture Techniques
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