Single Cell and Spheroid Collagen Type I Invasion Assay

Wever, Olivier De; Hendrix, An; Boeck, Astrid De; Eertmans, Frank; Westbroek, Wendy; Braems, Geert; Bracke, Marc

doi:10.1007/978-1-4614-8244-4_2

Cited by 14 publications

(18 citation statements)

References 19 publications

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“…This inhibits downstream intercellular signalling of the receptor [43]. The VEGF, FGF and PDGF family proteins are abundantly present in the tumourenvironment as they are secreted as well by cancer cells, CAFs and tumour-associated macrophages (TAMs) [21,22,24,60]. The receptors are expressed on the cell membrane of cancer cells, CAFs, TAMs and endothelial cells.…”

Section: Cafs As a Target In Cancer Therapymentioning

confidence: 99%

“…This shows that CAFs stimulate cancer cell invasion without the need for direct heterotypic cell-cell contacts or track formation. [22]. Soluble factors can be added in a gradient by placing the type I collagen gel on top of a transwell filter and the soluble factors of interest in the lower compartment [26] (Fig.…”

Section: Three-dimensional Biomimetic Approaches To Better Understandmentioning

confidence: 99%

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Cancer-associated fibroblasts as target and tool in cancer therapeutics and diagnostics

et al. 2015

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Cancer-associated fibroblasts (CAFs) are drivers of tumour progression and are considered as a target and a tool in cancer diagnostic and therapeutic applications. An increased abundance of CAFs or CAF signatures are recognized as a bad prognostic marker in several cancer types. Tumour-environment biomimetics strongly improve our understanding of the communication between CAFs, cancer cells and other host cells. Several experimental drugs targeting CAFs are in clinical trials for multiple tumour entities; alternatively, CAFs can be exploited as a tool to characterize the functionality of circulating tumour cells or to capture them as a tool to prevent metastasis. The continuous interaction between tissue engineers, biomaterial experts and cancer researchers creates the possibility to biomimic the tumour-environment and provides new opportunities in cancer diagnostics and management.

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Section: Cafs As a Target In Cancer Therapymentioning

confidence: 99%

Section: Three-dimensional Biomimetic Approaches To Better Understandmentioning

confidence: 99%

Cancer-associated fibroblasts as target and tool in cancer therapeutics and diagnostics

et al. 2015

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“…Cell invasion ability was observed by spheroid collagen type I invasion assay and tested by Transwell invasion assay based on published protocol [29,30]. For spheroid collagen type I invasion assay, HUVECs at a total number of 5.0 × 10 4 in 100 μL EGM-2 medium (Lonza) were seeded on 96-well ultra-low attachment surface plates (Corning) for 2 days to generate cell spheres.…”

Section: Cell Invasion Assaymentioning

confidence: 99%

SNAI1, an endothelial–mesenchymal transition transcription factor, promotes the early phase of ocular neovascularization

Sun

Chang

et al. 2018

Angiogenesis

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Ocular neovascularization is a comprehensive process involved in retinal vascular development and several blinding diseases such as age-related macular degeneration and retinopathy of prematurity, with vascular endothelial growth factor (VEGF) regarded as the master regulator. However, the qualified effect of anti-VEGF therapy reveals that the underlying mechanisms are still not clearly identified. To initialize angiogenesis, endothelial cells undergo a phenotype switching to generate highly migratory and invasive cells. This process shares certain similar characters observed in endothelial-mesenchymal transition (EndMT). Here, we found that SNAI1, an EndMT transcription factor, was expressed by endothelial cells in both physiological and pathological ocular neovascularization. SNAI1 overexpression triggered cell morphological change and enhanced cell motility, while loss of SNAI1 attenuated migration, invasion and sprouting. RNA sequence analysis further revealed that SNAI1 knockdown decreased the expression of genes related to cytoskeleton rearrangement and ECM remodeling. Moreover, intravitreal injection of small interfering RNA of SNAI1 suppressed new vessel formation in developing retina as well as mice model of choroidal neovascularization and oxygen-induced retinopathy. Therefore, we propose that the EndMT transcription factor SNAI1 promotes the early phase of ocular neovascularization and may provide a potential therapeutic target.

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“…Select spheroidal fragments exhibiting a thin, homogeneous layer of fibroblastic cells (forming a translucent capsule) and a diameter of 0. 4 mm by means of a macroscope and needles. One chick heart will yield about 20 suitable PHFs.…”

Section: Preparation Of Precultured Heart Fragments (Phfs)mentioning

confidence: 99%

“…This aspect is more complete than in other invasion assays in vitro that are based on tumor cell invasion into non-living gels composed of basement membrane 3 or interstitial matrix 4 molecules. The 11,12 .…”

Section: Introductionmentioning

confidence: 99%

Chick Heart Invasion Assay for Testing the Invasiveness of Cancer Cells and the Activity of Potentially Anti-invasive Compounds

Bracke¹,

Roman

Stevens

et al. 2015

JoVE

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The goal of the chick heart assay is to offer a relevant organ culture method to study tumor invasion in three dimensions. The assay can distinguish between invasive and non-invasive cells, and enables study of the effects of test compounds on tumor invasion. Cancer cells -either as aggregates or single cells -are confronted with fragments of embryonic chick heart. After organ culture in suspension for a few days or weeks the confronting cultures are fixed and embedded in paraffin for histological analysis. The three-dimensional interaction between the cancer cells and the normal tissue is then reconstructed from serial sections stained with hematoxylin-eosin or after immunohistochemical staining for epitopes in the heart tissue or the confronting cancer cells. The assay is consistent with the recent concept that cancer invasion is the result of molecular interactions between the cancer cells and their neighbouring stromal host elements (myofibroblasts, endothelial cells, extracellular matrix components, etc.). Here, this stromal environment is offered to the cancer cells as a living tissue fragment. Supporting aspects to the relevance of the assay are multiple. Invasion in the assay is in accordance with the criteria of cancer invasion: progressive occupation and replacement in time and space of the host tissue, and invasiveness and non-invasiveness in vivo of the confronting cells generally correlates with the outcome of the assay. Furthermore, the invasion pattern of cells in vivo, as defined by pathologists, is reflected in the histological images in the assay. Quantitative structure-activity relation (QSAR) analysis of the results obtained with numerous potentially anti-invasive organic congener compounds allowed the study of structure-activity relations for flavonoids and chalcones, and known anti-metastatic drugs used in the clinic (e.g., microtubule inhibitors) inhibit invasion in the assay as well. However, the assay does not take into account immunological contributions to cancer invasion. Video LinkThe video component of this article can be found at

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Single Cell and Spheroid Collagen Type I Invasion Assay

Cited by 14 publications

References 19 publications

Cancer-associated fibroblasts as target and tool in cancer therapeutics and diagnostics

Cancer-associated fibroblasts as target and tool in cancer therapeutics and diagnostics

SNAI1, an endothelial–mesenchymal transition transcription factor, promotes the early phase of ocular neovascularization

Chick Heart Invasion Assay for Testing the Invasiveness of Cancer Cells and the Activity of Potentially Anti-invasive Compounds

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