Desenvolvimento de modelos 3D de cancro da mama para avaliação de agentes anticancerígenos

  • Ana Rita Caldas Martins (Student)

Student thesis: Master's Thesis


The in vitro two-dimensional (2D) culture of cancer cells does not represent the three-dimensional (3D) architecture, heterogeneity, and complexity of human tumors. In this sense, more representative models, i.e., that better reflect the main aspects of tumor biology. These are essential studies in cancer biology and immunology, as well as for target validation and drug discovery. Three-dimensional (3D) cell culture systems have gained increasing interest for new drug discovery and tissue engineering given their clear advantages in providing more relevant physiologically information and more predictive data for testing in vivo. This dissertation discusses the characteristics of 3D cell culture systems compared to 2D culture. The main 3D cell culture technologies and their impact on drug discovery are also discussed, including the formation of spheroids using different techniques. Spheroids are the result of the growth and aggregation of one or more cell types in 3D culture. In general, spheroids retain the characteristics of the initial cells but assume a spherical shape. This work consists of an optimization process for the formation of spheroids from MDA-MB-231 cells, which is a triple-negative breast cancer (CMTN) cell line, through the liquid overlay culture technique that includes U-bottom plates and the use of agarose as a substrate. The response of MDA-MB-231 cells to Doxorubicin (DOX) was evaluated in the 3D cell culture model and compared to a response using the monolayer (2D) culture model. Through a quantitative analysis based on the growing area of the spheroids, it was found that the 3D cultures increase the susceptibility to DOX at the concentration of 4 μg/mL, in which a greater reduction in the spheroid area was observed. Exposure to DOX led to a greater decrease in the number of viable cells in the 2D culture model compared to the 3D culture model, demonstrating that is more resistant to chemotherapeutics. In summary, this work studies the formation, structure, and growth of spheroids, as well as their toxicological drug resistance compared to cell culture in a 2D model.
Date of Award3 Mar 2022
Original languagePortuguese
Awarding Institution
  • Universidade Católica Portuguesa
SupervisorLígia R. Rodrigues (Supervisor)


  • Triple-negative breast cancer
  • 2D culture
  • 3D culture
  • Spheroids
  • Doxorubicin


  • Mestrado em Engenharia Biomédica

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