Vascular Endothelial–Breast Epithelial Cell Coculture Model Created from 3D Cell Structures

Swaminathan, Swathi; Ngo, Olivia; Basehore, Sarah; Clyne, Alisa Morss

doi:10.1021/acsbiomaterials.6b00624

Cited by 10 publications

(12 citation statements)

References 30 publications

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“…In our previous study, we showed that MCF10A-NeuN breast epithelial cells migrated out of spheroids and along HUVEC tubes as soon as they were placed in coculture. 9 AD-01 treatment significantly inhibited MCF10A-NeuN spheroid adhesion to and migration along HUVEC tube-like networks more so than for MCF10A spheroids. ERBB2/HER2 is regarded as a poor prognostic indicator for breast cancer survival, and its overexpression drives tumor cell survival, tumor growth, and metastasis.…”

Section: Discussionmentioning

confidence: 82%

“…We formed breast cell 3D spheroids using an established protocol. 9,34 Briefly, we added 30 mL undiluted Matrigel (Corning; growth factor reduced) to each well of a BioCoat Falcon Culture slide (Corning), which was then gelled for 20 min at 37°C. Approximately, 5000 breast epithelial cells were seeded in each well in the appropriate culture medium with 20% added Matrigel.…”

Section: Formation Of Breast Cell 3d Spheroidsmentioning

confidence: 99%

“…We previously observed that breast 3D spheroids attached to and migrated along HUVEC tube-like networks in coculture. 9 We therefore investigated the AD-01 effect on breast spheroid adhesion to HUVEC networks. AD-01 did not affect HUVEC tube-like networks over 48 h of treatment (Fig.…”

Section: Ad-01 Decreased 2d and 3d Breast Epithelial Cell Adhesionmentioning

confidence: 99%

“…[1][2][3][4] Three-dimensional (3D) human cancer models, such as breast epithelial spheroids, better recapitulate many in vivo cell functions, including increased chemoresistance. [4][5][6][7][8] More recently, 3D culture models were designed to also include other cell types through direct contact 9,10 or Transwell plates. 11,12 These 3D coculture models enhance the cell-cell interactions that are necessary to study adhesion, migration, and tumor invasion.…”

Section: Introductionmentioning

confidence: 99%

“…7,[13][14][15] We recently created a 3D coculture model system of endothelial cell tube-like networks and breast epithelial spheroids, which demonstrated that breast epithelial spheroids preferentially adhered to and migrated along the endothelial networks. 9 CD44 is a cell surface adhesion molecule that is upregulated in breast cancer and is gaining importance in identifying cancer stem cells and as a cancer diagnostic marker. 16,17 CD44 expression is thought to negatively correlate with breast cancer disease-free survival in human patients and, in a mouse model, CD44 knockdown reduced MDA-MB-231 tumor burden and increased survival.…”

Section: Introductionmentioning

confidence: 99%

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A Three-Dimensional In Vitro Coculture Model to Quantify Breast Epithelial Cell Adhesion to Endothelial Cells

Swaminathan

Cranston

Clyne

2019

Tissue Engineering Part C: Methods

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Three-dimensional (3D) in vitro culture models better recapitulate the tissue microenvironment, and therefore may provide a better platform to evaluate therapeutic effects on adhesive cell-cell interactions. The objective of this study was to determine if AD-01, a peptide derivative of FK506-binding protein like that is reported to bind to the adhesion receptor CD44, would induce a greater reduction in breast epithelial spheroid adhesion to endothelial tube-like networks in our 3D coculture model system compared to two-dimensional (2D) culture. MCF10A, MCF10A-NeuN, MDA-MB-231, and MCF7 breast epithelial cells were pretreated with AD-01 either as single cells or as spheroids. Breast epithelial cell adhesion to 2D tissue culture substrates was first measured, followed by spheroid formation (breast cell-cell adhesion) and spheroid adhesion to Matrigel or endothelial networks. Finally, CD44 expression was quantified in breast epithelial cells in 2D and 3D culture. Our results show that AD-01 had the largest effect on spheroid formation, specifically in breast cancer cell lines. AD-01 also inhibited breast cancer spheroid adhesion to and migration along endothelial networks. The different breast epithelial cell lines expressed more CD44 when cultured as 3D spheroids, but this did not universally translate into higher protein levels. This study shows that 3D coculture models can enable unique insights into cell adhesion, migration, and cell-cell interactions, thereby enhancing understanding of basic biological mechanisms. Furthermore, such 3D coculture systems may also represent a more relevant testing platform for understanding the mechanism-of-action of new therapeutic agents.

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Section: Discussionmentioning

confidence: 82%

Section: Formation Of Breast Cell 3d Spheroidsmentioning

confidence: 99%

Section: Ad-01 Decreased 2d and 3d Breast Epithelial Cell Adhesionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Three-Dimensional In Vitro Coculture Model to Quantify Breast Epithelial Cell Adhesion to Endothelial Cells

Swaminathan

Cranston

Clyne

2019

Tissue Engineering Part C: Methods

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Targeting resistant breast cancer stem cells in a three-dimensional culture model with oleuropein encapsulated in methacrylated alginate microparticles

Altundag-Erdogan,

Tutar,

Yüce

et al. 2024

DARU J Pharm Sci

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Background Cancer stem cells (CSCs) are a subpopulation of cancer cells that are believed to be responsible for tumor initiation, progression, metastasis, and resistance to conventional therapies. Oleuropein as a natural compound found in olive leaves and olive oil, has potential therapeutic effects in cancer treatment, particularly in targeting CSCs. It induces apoptosis in CSCs while sparing normal cells, inhibit proliferation, migration, and invasion, and suppress the self-renewal ability of CSCs. Additionally, oleuropein has shown synergistic effects with conventional chemotherapy drugs, enhancing their efficacy against CSCs. Objectives This study aims to selectively target therapeutically resistant cancer stem cells (CSCs) within a heterogeneous tumor population by utilizing oleuropein (OLE) encapsulated in methacrylated alginate (OLE-mALG) within an in vivo-like microenvironment. Purpose This study aims to target therapeutically resistant cancer stem cells (CSCs) with oleuropein (OLE) encapsulated in the methacrylated alginate (OLE-mALG) in a heterogeneous tumor population with an in vivo-like microenvironment. Methods Co-culture of CSCs with non-tumorogenic MCF-12 A cells was performed, the 3D breast cancer model was supported with methocel/matrigel/collagen-I, and vascularization was ensured with human umbilical vein endothelial cells (HUVEC). Then, OLE-loaded methacrylated alginate microparticles (mALG) were formed by dual crosslinking in the presence of both ionic and visible light obtained with a droplet based microfluidic system. The characterization and effectiveness of the produced OLE-mALG were evaluated by the FTIR, swelling/degradation/release analysis. Before producing OLE loaded mALG microparticles, a preliminary study was carried out to determine the effective dose of OLE for cells and the duration of OLE action on MCF-7, CSCs and MCF-12 A. Subsequently, CSC viability (WST-1), apoptosis (Bcl-2, Bax, caspase-3, caspase-9), stemness (OCT3/4, NANOG, SOX2), EMT profile (E-cadherin, Vimentin, Slug) and proliferation (SURVIVIN, p21, CYCLIN D1) after OLE-mALG treatment were all evaluated in the 3D model. Results OLE was encapsulated in mALG with an efficiency of 90.49% and released 73% within 7 h. OLE-mALG induced apoptosis through the decrease in anti-apoptotic Bcl-2 and an increase in pro-apoptotic Bax, caspase-3, and caspase-9 protein levels. While Vimentin and Slug protein levels decreased after 200 µg/mL OLE-mALG treatment to 3D breast cancer culture, E-cadherin levels increased. OLE-mALG treatment to CSC co-culture led to a decrease in proliferation by triggering p21/SURVIVIN expressions, and also resulted in an increase in stemness genes (OCT3/4/NANOG/SOX2). Conclusion 200 µg/mL OLE-loaded mALG microparticles suppressed epithelial-to-mesenchymal transition by suppressing Vimentin and Slug protein levels, and increased E-cadherin levels in the 3D breast cancer model we created with CSCs, MCF-12 A and HUVECs. This complex system may allow the use of personalized cells for rapid drug screening in preclinical studies compared to animal experiments. OLE-mALG showed apoptotic and metastasis suppressive properties in cancer cells and it was concluded that it can be used in combination with or alternatively with chemotherapeutic agents to target breast cancer stem cells. Graphical abstract

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Multi compartmental 3D breast cancer disease model–recapitulating tumor complexity in in-vitro

Nair

Mukherjee

Kaur

et al. 2023

Biochimica et Biophysica Acta (BBA) - General Subjects

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Vascular Endothelial–Breast Epithelial Cell Coculture Model Created from 3D Cell Structures

Cited by 10 publications

References 30 publications

A Three-Dimensional In Vitro Coculture Model to Quantify Breast Epithelial Cell Adhesion to Endothelial Cells

A Three-Dimensional In Vitro Coculture Model to Quantify Breast Epithelial Cell Adhesion to Endothelial Cells

Targeting resistant breast cancer stem cells in a three-dimensional culture model with oleuropein encapsulated in methacrylated alginate microparticles

Multi compartmental 3D breast cancer disease model–recapitulating tumor complexity in in-vitro

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