Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures

Triple-negative breast cancer (TNBC) is highly aggressive and lacks targeted therapies, posing a major challenge in oncology. Traditional two-dimensional (2D) cell cultures fail to capture the tumor microenvironment’s complexity, whereas three-dimensional (3D) cultures provide a more accurate model...

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Main Authors: Mercedes Olvera-Valencia, Verónica Garcia-Castillo, Rosalío Ramos-Payan, Maribel Aguilar-Medina, Samuel Trujano-Camacho, Alejandro López-Saavedra, Laurence A. Marchat, Cesar López-Camarillo, Ronen Sumagin, Eloy Pérez-Yepez, Carlos Pérez-Plasencia
Format: Article
Language:English
Published: SAGE Publishing 2025-05-01
Series:Journal of Tissue Engineering
Online Access:https://doi.org/10.1177/20417314251326668
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author Mercedes Olvera-Valencia
Verónica Garcia-Castillo
Rosalío Ramos-Payan
Maribel Aguilar-Medina
Samuel Trujano-Camacho
Alejandro López-Saavedra
Laurence A. Marchat
Cesar López-Camarillo
Ronen Sumagin
Eloy Pérez-Yepez
Carlos Pérez-Plasencia
author_facet Mercedes Olvera-Valencia
Verónica Garcia-Castillo
Rosalío Ramos-Payan
Maribel Aguilar-Medina
Samuel Trujano-Camacho
Alejandro López-Saavedra
Laurence A. Marchat
Cesar López-Camarillo
Ronen Sumagin
Eloy Pérez-Yepez
Carlos Pérez-Plasencia
author_sort Mercedes Olvera-Valencia
collection DOAJ
description Triple-negative breast cancer (TNBC) is highly aggressive and lacks targeted therapies, posing a major challenge in oncology. Traditional two-dimensional (2D) cell cultures fail to capture the tumor microenvironment’s complexity, whereas three-dimensional (3D) cultures provide a more accurate model of tumor biology. We developed an advanced 3D culture system for TNBC cell lines BT-20 and MDA-MB-231, enhancing the hanging-drop method with Matrigel to restore essential extracellular matrix interactions. Confocal imaging showed MDA-MB-231 cells forming clusters typical of aggressive carcinoma, while BT-20 cells organized into duct-like structures. Molecular analysis of PI3K and β-catenin target genes revealed distinct expression patterns, with PI3K overexpressed and β-catenin downregulated in 3D cultures. Moreover, β-catenin distribution in the 3D cell culture closely resembles its pattern in tissue. These findings underscore the value of 3D models in understanding TNBC progression and in supporting the exploration of novel therapeutic strategies.
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spelling doaj-art-4aede081820243fda28d6824195acce12025-08-20T02:57:17ZengSAGE PublishingJournal of Tissue Engineering2041-73142025-05-011610.1177/20417314251326668Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D culturesMercedes Olvera-Valencia0Verónica Garcia-Castillo1Rosalío Ramos-Payan2Maribel Aguilar-Medina3Samuel Trujano-Camacho4Alejandro López-Saavedra5Laurence A. Marchat6Cesar López-Camarillo7Ronen Sumagin8Eloy Pérez-Yepez9Carlos Pérez-Plasencia10Laboratorio de Genómica, Instituto Nacional de Cancerología, Tlalpan, CDMX, MexicoLaboratorio de Genómica Funcional, Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, UNAM, Tlalnepantla Estado de México, MexicoFacultad de Ciencias Químico Biológicas, Universidad Autónoma de Sinaloa, Culiacan, Sinaloa, MexicoFacultad de Ciencias Químico Biológicas, Universidad Autónoma de Sinaloa, Culiacan, Sinaloa, MexicoExperimental Biology PhD Program, DCBS, Universidad Autónoma Metropolitana- Iztapalapa, Iztapalapa, MexicoTecnológico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Mexico City, MexicoPrograma Institucional de Biomedicina Molecular, Escuela Nacional de Medicina y Homeopatía del Instituto Politécnico Nacional, Ticoman, CDMX, MexicoPosgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, Benito Juarez, CDMX, MexicoDepartment of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USALaboratorio de Genómica, Instituto Nacional de Cancerología, Tlalpan, CDMX, MexicoLaboratorio de Genómica Funcional, Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, UNAM, Tlalnepantla Estado de México, MexicoTriple-negative breast cancer (TNBC) is highly aggressive and lacks targeted therapies, posing a major challenge in oncology. Traditional two-dimensional (2D) cell cultures fail to capture the tumor microenvironment’s complexity, whereas three-dimensional (3D) cultures provide a more accurate model of tumor biology. We developed an advanced 3D culture system for TNBC cell lines BT-20 and MDA-MB-231, enhancing the hanging-drop method with Matrigel to restore essential extracellular matrix interactions. Confocal imaging showed MDA-MB-231 cells forming clusters typical of aggressive carcinoma, while BT-20 cells organized into duct-like structures. Molecular analysis of PI3K and β-catenin target genes revealed distinct expression patterns, with PI3K overexpressed and β-catenin downregulated in 3D cultures. Moreover, β-catenin distribution in the 3D cell culture closely resembles its pattern in tissue. These findings underscore the value of 3D models in understanding TNBC progression and in supporting the exploration of novel therapeutic strategies.https://doi.org/10.1177/20417314251326668
spellingShingle Mercedes Olvera-Valencia
Verónica Garcia-Castillo
Rosalío Ramos-Payan
Maribel Aguilar-Medina
Samuel Trujano-Camacho
Alejandro López-Saavedra
Laurence A. Marchat
Cesar López-Camarillo
Ronen Sumagin
Eloy Pérez-Yepez
Carlos Pérez-Plasencia
Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures
Journal of Tissue Engineering
title Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures
title_full Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures
title_fullStr Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures
title_full_unstemmed Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures
title_short Development of a reliable method for human triple-negative breast cancer organotypic culture: Improving imaging and genomic studies in 3D cultures
title_sort development of a reliable method for human triple negative breast cancer organotypic culture improving imaging and genomic studies in 3d cultures
url https://doi.org/10.1177/20417314251326668
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