MCF7 Spheroid Development: New Insight about Spatio/Temporal Arrangements of TNTs, Amyloid Fibrils, Cell Connections, and Cellular Bridges

Pulze, Laura; Congiu, Terenzio; Brevini, Tiziana A. L.; Grimaldi, Annalisa; Tettamanti, Gianluca; D’Antona, Paola; Baranzini, Nicolò; Acquati, Francesco; Ferraro, Federico; Eguileor, Magda de

doi:10.3390/ijms21155400

Cited by 22 publications

(17 citation statements)

References 80 publications

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“…Detailed structure of the spheroids (especially seen on the microscope computer screen) suggest that the spheroids are mostly free to move in the medium. This agrees with a publication [ 19 ]. Our protocol for producing and maintaining tumoroids can be used for cancer drug screening, discovery, and development.…”

Section: Resultssupporting

confidence: 94%

“…MCF-7 is a commonly used breast cancer cell line for breast cancer research for more than 40 years by multiple research groups, it is an estrogen-dependent breast cancer cell line [ 18 ]. Recently, MCF-7 cells have been used to develop tumoroids in ultra-low attachment 6-well plates in culture medium [ 19 ], resulting in new insights about spatio-temporal arrangements of tunneling nanotubes, amyloid fibrils, cell connections, and cellular bridges. We have recently developed a simple, inexpensive method for MCF-7 cell spheroid growth in 96-well low attachment plates.…”

Section: Introductionmentioning

confidence: 99%

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Breast Cancer MCF-7 Cell Spheroid Culture for Drug Discovery and Development

Chen¹,

Liu²,

Yan³

2022

JCT

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In vitro 3D cancer spheroids (tumoroids) exhibit a drug resistance profile similar to that found in solid tumors. 3D spheroid culture methods recreate more physiologically relevant microenvironments for cells. Therefore, these models are more appropriate for cancer drug screening. We have recently developed a protocol for MCF-7 cell spheroid culture, and used this method to test the effects of different types of drugs on this estrogen-dependent breast cancer cell spheroid. Our results demonstrated that MCF-7 cells can grow spheroid in medium using a low attachment plate. We managed to grow one spheroid in each well, and the spheroid can grow over a month, the size of the spheroid can grow over a hundred times in volume. Our targeted drug experimental results suggest that estrogen sulfotransferase, steroid sulfatase, and G protein-coupled estrogen receptor may play critical roles in MCF-7 cell spheroid growth, while estrogen receptors α and β may not play an essential role in MCF-7 spheroid growth. Organoids are the miniatures of in vivo tissues and reiterate the in vivo microenvironment of a specific organ, best fit for the in vitro studies of diseases and drug development. Tumoroid, developed from cancer cell lines or patients’ tumor tissue, is the best in vitro model of in vivo tumors. 3D spheroid technology will be the best future method for drug development of cancers and other diseases. Our reported method can be developed clinically to develop personalized drugs when the patient’s tumor tissues are used to develop a spheroid culture for drug screening.

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Section: Resultssupporting

confidence: 94%

Section: Introductionmentioning

confidence: 99%

Breast Cancer MCF-7 Cell Spheroid Culture for Drug Discovery and Development

Chen¹,

Liu²,

Yan³

2022

JCT

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“…While identifying straight membranous protrusions with no substratum contact is achievable in a sparse, 2D cell culture landscape, identifying TNTs in 3D models such as organoids (Zhang et al , 2018; Pulze et al , 2020; Whitehead et al , 2020), ex vivo explants (Chinnery et al , 2008), and tissues (Lou et al , 2012), is considerably more challenging. In these and/or other ecosystems in which cells are embedded in a natural or artificial matrix, stromal elements, such as neighboring cells, can both overshadow TNTs and force them to adopt convoluted shapes compared to those seen in vitro (Korenkova et al , 2020).…”

Section: An Overview Of Tunneling Nanotubesmentioning

confidence: 99%

“…While identifying straight membranous protrusions with no substratum contact is achievable in a sparse, 2D cell culture landscape, identifying TNTs in 3D models such as organoids (Zhang et al, 2018;Pulze et al, 2020;Whitehead et al, 2020), ex vivo explants (Chinnery et al, 2008), and tissues (Lou et al, 2012), is considerably more challenging. In these and/or other ecosystems in TNTs are (I) thin (20-700 nm) and long reaching (up to 100 µm in thin TNTs; < 700 nm in diameter) membranous protrusions that hover over the substrate and connect distant cells.…”

Section: An Overview Of Tunneling Nanotubesmentioning

confidence: 99%

Peering into tunneling nanotubes—The path forward

Cervantes

Zurzolo

2021

The EMBO Journal

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The identification of Tunneling Nanotubes (TNTs) and TNT‐like structures signified a critical turning point in the field of cell‐cell communication. With hypothesized roles in development and disease progression, TNTs’ ability to transport biological cargo between distant cells has elevated these structures to a unique and privileged position among other mechanisms of intercellular communication. However, the field faces numerous challenges—some of the most pressing issues being the demonstration of TNTs in vivo and understanding how they form and function. Another stumbling block is represented by the vast disparity in structures classified as TNTs. In order to address this ambiguity, we propose a clear nomenclature and provide a comprehensive overview of the existing knowledge concerning TNTs. We also discuss their structure, formation‐related pathways, biological function, as well as their proposed role in disease. Furthermore, we pinpoint gaps and dichotomies found across the field and highlight unexplored research avenues. Lastly, we review the methods employed to date and suggest the application of new technologies to better understand these elusive biological structures.

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“…The ability of cells to self-assemble is not extraordinary itself, but usual to culturing the cells in 3D models a special systems or supplements, e.g., hanging drop, scaffolds and hydrogels, special plastic, and cocktails of growth factors are needed [ 2 , 23 ]. Various methods have been described for obtaining 3D MCF7 cell culture [ 20 , 24 , 25 , 26 ]. Depending on the technology used, the basic molecular characteristics of the cells in 3D may vary, which can result in difficulties in reproducing such models.…”

Section: Discussionmentioning

confidence: 99%

EGFR Transgene Stimulates Spontaneous Formation of MCF7 Breast Cancer Cells Spheroids with Partly Loss of HER3 Receptor

Troitskaya

Novak

Nushtaeva

et al. 2021

IJMS

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Multicellular spheroids with 3D cell–cell interactions are a useful model to simulate the growth conditions of cancer. There is evidence that in tumor spheroids, the expression of various essential molecules is changed compared to the adherent form of cell cultures. These changes include growth factor receptors and ABC transporters and result in the enhanced invasiveness of the cells and drug resistance. It is known that breast adenocarcinoma MCF7 cells can spontaneously form 3D spheroids and such spheroids are characterized by high expression of EGFR/HER2, while the natural phenotype of MCF7 cells is EGFRlow/HER2low. Therefore, it was interesting to reveal if high epidermal growth factor receptor (EGFR) expression is sufficient for the conversion of adherent MCF7 to spheroids. In this study, an MCF7 cell line with high expression of EGFR was engineered using the retroviral transduction method. These MCF7-EGFR cells assembled in spheroids very quickly and grew predominantly as a 3D suspension culture with no special plates, scaffolds, growth supplements, or exogenous matrixes. These spheroids were characterized by a rounded shape with a well-defined external border and 100 µM median diameter. The sphere-forming ability of MCF7-EGFR cells was up to 5 times stronger than in MCF7wt cells. Thus, high EGFR expression was the initiation factor of conversion of adherent MCF7wt cells to spheroids. MCF7-EGFR spheroids were enriched by the cells with a cancer stem cell (CSC) phenotype CD24−/low/CD44− in comparison with parental MCF7wt cells and MCF7-EGFR adhesive cells. We suppose that these properties of MCF7-EGFR spheroids originate from the typical features of parental MCF7 cells. We showed the decreasing of HER3 receptors in MCF7-EGFR spheroids compared to that in MCFwt and in adherent MCF7-EGFR cells, and the same decrease was observed in the MCF7wt spheroids growing under the growth factors stimulation. To summarize, the expression of EGFR transgene in MCF7 cells stimulates rapid spheroids formation; these spheroids are enriched by CSC-like CD24−/CD44− cells, they partly lose HER3 receptors, and are characterized by a lower potency in drug resistance pomp activation compared to MCF7wt. These MCF7-EGFR spheroids are a useful cancer model for the development of anticancer drugs, including EGFR-targeted therapeutics.

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MCF7 Spheroid Development: New Insight about Spatio/Temporal Arrangements of TNTs, Amyloid Fibrils, Cell Connections, and Cellular Bridges

Cited by 22 publications

References 80 publications

Breast Cancer MCF-7 Cell Spheroid Culture for Drug Discovery and Development

Breast Cancer MCF-7 Cell Spheroid Culture for Drug Discovery and Development

Peering into tunneling nanotubes—The path forward

EGFR Transgene Stimulates Spontaneous Formation of MCF7 Breast Cancer Cells Spheroids with Partly Loss of HER3 Receptor

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