Morphometric analysis of a triple negative breast cancer cell line in hydrogel and monolayer culture environments

Area of Science:

• Biomedical Engineering
• Cell Biology
• Oncology

Background:

• Triple negative breast cancer (TNBC) lacks targeted therapies, necessitating novel treatment strategies.
• Advanced in vitro cell culture models are crucial for understanding TNBC and identifying new therapeutic targets.
• Three-dimensional (3D) culture systems, particularly those using hydrogel scaffolds, can better mimic the tumor microenvironment.

Purpose of the Study:

• To establish and evaluate protocols for culturing the TNBC cell line HCC70 in both monolayer and 3D hydrogel environments.
• To compare the morphology and subcellular organization of TNBC cells grown under different culture conditions.
• To provide insights into TNBC cell behavior and adaptation within varying in vitro microenvironments.

Main Methods:

• Cultured HCC70 cells in both standard monolayer and basement membrane matrix hydrogel conditions.
• Utilized light microscopy (phase contrast, confocal) for morphological assessment.
• Employed transmission electron microscopy (TEM) for quantitative ultrastructural analysis of subcellular components.

Main Results:

• Monolayer cultures exhibited flattened, irregularly shaped cells.
• Hydrogel cultures promoted the formation of multi-layered cell spheroids.
• Spheroid cells showed a higher abundance of mitochondria, autophagic vacuoles, and intercellular junctions compared to monolayer cells.

Conclusions:

• Triple negative breast cancer cells exhibit altered morphology and organelle content in response to their microenvironment.
• 3D hydrogel culture systems provide a more biologically relevant in vitro model for TNBC compared to traditional monolayers.
• These findings support the use of diverse in vitro models, including 3D hydrogels, for advancing TNBC research and therapeutic target discovery.