Abstract
Triple negative breast cancer (TNBC) is a belligerent carcinoma that is unresponsive to targeted receptor therapies. Development of new treatment strategies would benefit from an expanded repertoire of in vitro cell culture systems, such as those that support tridimensional growth in the presence of hydrogel scaffolds. To this end, we established protocols for maintenance of the TNBC cell line HCC70 in monolayer culture and in a commercially available basement membrane matrix hydrogel. We evaluated the general morphology of cells grown in both conditions with light microscopy, and examined their subcellular organization using transmission electron microscopy (TEM). Phase contrast and confocal microscopy showed the prevalence of irregularly shaped flattened cells in monolayer cultures, while cells maintained in hydrogel organized into multi-layered spheroids. A quantitative ultrastructural analysis comparing cells from the two culture conditions revealed that cells that formed spheroids comprised a greater number of mitochondria, autophagic vacuoles and intercellular junctions than their monolayer counterparts, within the equivalent area of sampled tissue. These observations suggest that triple negative breast cancer cells in culture can alter their organelle content, as well as their morphology, in response to their microenvironment. Methods presented here may be useful for those who intend to image cell cultures with TEM, and for investigators who seek to implement diverse in vitro models in the search for therapeutic molecular targets for TNBC.
Highlights
Triple negative breast cancer (TNBC) is an invasive cancer that spreads rapidly, weakens the body and can cause discomfort and pain
Phase contrast and confocal microscopy of monolayer and hydrogel cultures The general morphology of HCC70 cells was examined using phase contrast microscopy (Fig. 2)
Confocal images of HCC70 cells in monolayer cultures indicated that cells were flattened and adhered to the rigid glass substratum, while cells cultured in the hydrogel formed well-rounded, multilayered structures
Summary
Triple negative breast cancer (TNBC) is an invasive cancer that spreads rapidly, weakens the body and can cause discomfort and pain. Mutations in BRCA genes further increase the susceptibility of women to this disease (Meyer et al, 2012; Kuo. How to cite this article Jogalekar and Serrano (2018), Morphometric analysis of a triple negative breast cancer cell line in hydrogel and monolayer culture environments. Poor prognosis and a high recurrence have been linked with TNBC resistance to chemotherapy and radiation therapy (Dent et al, 2007; Zhang et al, 2008; Atkinson et al, 2010; Boyle, 2012). Some reports suggest that an increase in cancer stem cells and autophagy may play a major role in imparting chemo- and radio-resistance to TNBC cells (Atkinson et al, 2013; O’Reilly et al, 2015). The mandate for new treatments against TNBC is spurring development of innovative approaches, such as the use of iron oxide nanoparticles and temperature-sensitive liposomes conjugated with therapeutic drugs (Guo et al, 2014; Ou et al, 2016)
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