Cited by 9343D Printing in Biomedical ResearchCancer Cells and MetastasisMathematical Biology Tumor Growth

Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types

Kelm, Jens M., Timmins, Nicholas E., Brown, Catherine J., Fussenegger, Martin, Nielsen, Lars K.·2003·Biotechnology and Bioengineering

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.

Integrity signals

Registry

Crossref · 10.1002/bit.10655

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Cites

Flagged first, then by citations

Cited by

Sorted by citations

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

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review