3D organoid culture of MMTV-PyMT mammary gland tumors

Commonly used acronym: PyMT organoids

Scope of the method

The Method relates to
  • Animal health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
Type of cells/tissues/organs
mammary gland tumor


Method keywords
  • mammary gland tumors
  • carcinoma
  • mammary tumor organoids
  • Organoid model
Scientific area keywords
  • breast cancer
  • tumor-derived
  • 3D culture
Method description

The mammary-specific polyomavirus middle T antigen overexpression mouse model (MMTV-PyMT) is one of the most commonly used models in the cancer research field for multiple reasons, among which the spontaneous development of multifocal luminal tumors, the early tumoral onset, and the primary tumors' morphology resembling those in clinical biopsies. Here we present a method for the derivation of MMTV-PyMT tumor organoids recapitulating the complex structural organization and heterogeneity observed in vivo. The generation of an organotypic culture from MMTV-PyMT primary tumors provides a valid research platform exploitable for a variety of experimental analyses concerning the study of breast cancer progression, the modulation of the tumor-microenvironment or metastases formation. These organoids can be derived from a limited amount of starting material and are easily maintained and expanded in basement membrane extract (BME).

Lab equipment
  • - Biosafety cabinet,
  • - Cell incubator,
  • - Phase-contrast microscope.

Pros, cons & Future potential

  • - Primary cells,
  • - 3D organoids mimic tumor heterogeneity,
  • - Organoids have strong expandability and ease of maintenance,
  • - Versatile applications.

Recapitulation of the primary tumor but not of other cell types in the microenvironment that may play a role in the tumorigenic transformation and/progression.


Co-culturing with other cell types.

References, associated documents and other information

Contact person

Beaunelle de Bruijn


Oncology - KU Leuven

Katholieke Universiteit Leuven (KUL)