Stem cell proliferation patterns as an alternative for in vivo prediction and discrimination of carcinogenic compounds

Scope of the method

The Method relates to
  • Animal health
  • Environment
  • Human health
The Method is situated in
  • Translational - Applied Research
Type of method
  • In vivo
Used species
Planarians (i.e. Schmidtea mediterranea)
Targeted organ system or type of research
Full organism model (stem cell proliferation)


Method keywords
  • Planarians
  • Stem cell proliferation patterns
  • in vivo
Scientific area keywords
  • Genotoxic carcinogen
  • Non-genotoxic carcinogen
  • genotoxicity
Method description

The method uses planarians as an alternative in vivo model to assess carcinogenicity. Planarians are characterized by a high regenerative capacity and a large number of pluripotent stem cells. The assay is based on the discriminative power of stem cells in an in vivo setting. Based on specific stem cell dynamics and proliferation patterns, this method predicts carcinogenic potential and discriminates between genotoxic and non-genotoxic compounds. The workflow includes: (1) Exposure of planarians to the compound of interest, (2) Fixation of the animals at specific time points, (3) Immunohistochemical staining of the proliferating stem cells, (4) Quantification of the number of proliferating stem cells, (5) Determination of the resulting patterns in proliferative responses.

Lab equipment
  • - Facility for planarian culture
  • - Shaker
  • - Incubator or oven
  • - Fluorescent microscope
Method status
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

  • - Reduced use of laboratory animals, the method can be used as an initial screening
  • - Inexpensive and time-effective method
  • - Fast learning curve to apply the method successfully
  • - Includes biological variation
  • - Stem cells are studied in their natural context in a full organism model, including all communication signals

Further validation with additional compounds is needed.


Possibility to upscale and work in a high-throughput setting.

Future & Other applications

Inclusion of additional parameters (e.g. phenotypes, gene-expression) to increase the discriminative power and to identify the mode of action in detail.

References, associated documents and other information

  • Stevens AS, Willems M, Plusquin M, Ploem JP, Winckelmans E, Artois T, Smeets K. Stem cell proliferation patterns as an alternative for in vivo prediction and discrimination of carcinogenic compounds. Sci Rep. 2017 May 3;7:45616. doi: 10.1038/srep45616. PubMed PMID: 28466856; PubMed Central PMCID: PMC5413882.
  • Willems, M., Stevens A.S., Adriaens E., Plusquin M., Smeets K., Van Goethem F., Vanparys P., Janssen C., Remon J.P. (2015) An adult stem cell proliferation assay in the flatworm model Macrostomum lignano to predict the carcinogenicity of compounds. Applied in vitro toxicology 1 (3), 213-219.
  • Patent granted: Methods to defect carcinogens using flatworms WO 2016/146620 /EP15159158.3 A1 – Karen Smeets, An-Sofie Stevens, Michelle Plusquin, Tom Artois, Maxime Willems, Jean Paul Remon- filing date 15/03/2016

Contact person

Karen Smeets


University of Hasselt (UHasselt)
Centre for Environmental Sciences
Zoology: Biodiversity and Toxicology (BiTE Lab)
Flemish Region