Micropatterns of Matrigel for three-dimensional epithelial cultures

Sodunke, Temitope R; Turner, Keneshia K; Caldwell, Sarah A.; McBride, Kevin; Reginato, Mauricio J.; Noh, Hyunwoo

doi:10.1016/j.biomaterials.2007.05.021

Cited by 90 publications

(62 citation statements)

References 31 publications

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“…29,30 If it seems complicated to regulate spatial proteic micropattern in Figure 7 Anticancer drug testing in egg white matrix. (a) Aorta sections from GFP-expressing rats were seeded in 24-well plates coated with egg white.…”

Section: Discussionmentioning

confidence: 99%

In vitro 3D angiogenesis assay in egg white matrix: comparison to Matrigel, compatibility to various species, and suitability for drug testing

Mousseau

Mollard

Qiu

et al. 2014

Laboratory Investigation

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In vitro angiogenesis assays are commonly used to assess pro-or anti-angiogenic drug properties. Extracellular matrix (ECM) substitutes such as Matrigel and collagen gel became very popular in in vitro 3D angiogenesis assays as they enable tubule formation by endothelial cells from culture or aortic rings. However, these assays are usually used with a single cell type, lacking the complex cellular interactions occurring during angiogenesis. Here, we report a novel angiogenesis assay using egg white as ECM substitute. We found that, similar to Matrigel, egg white elicited prevascular network formation by endothelial and/or smooth muscle cell coculture. This matrix was suitable for various cells from human, mouse, and rat origin. It is compatible with aortic ring assay and also enables vascular and tumor cell coculture. Through simple labeling (DAPI, Hoechst 33258), cell location and resulting prevascular network formation can easily be quantified. Cell transfection with green fluorescent protein improved whole cell visualization and 3D structure characterization. Finally, eggbased assay dedicated to angiogenesis studies represents a reliable and cost-effective way to produce and analyze data regarding drug effects on vascular cells.

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Section: Discussionmentioning

confidence: 99%

In vitro 3D angiogenesis assay in egg white matrix: comparison to Matrigel, compatibility to various species, and suitability for drug testing

Mousseau

Mollard

Qiu

et al. 2014

Laboratory Investigation

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“…A potential drawback of using this approach is that ECM is a biological substance and, therefore, its exact composition varies between batches. Also, the costs associated with purchasing the matrix must be taken into account when considering large-scale production of 3D spheroids for high-throughput drug testing [36]. Furthermore, spheroid sizes tend not to be uniform, which may be undesirable for reproducible high-throughput drug testing.…”

Section: Matricesmentioning

confidence: 99%

“…Perhaps the biggest issue with using ECM for this purpose is that cells tend to be unevenly distributed throughout the product. This can result in spheroids overlapping with each other while in culture, which is undesirable when single spheroid analysis is required [36].…”

Section: Matricesmentioning

confidence: 99%

Three-dimensional cell culture: the missing link in drug discovery

Breslin

O’Driscoll

2013

Drug Discovery Today

1,077

933

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“…Alternative cell culture systems are based on 3D cell culture techniques mimicking the microenvironment of tissues or organs and allow long term maintenance of the cells. These cultivation systems include gel-based systems imitating extracellular matrix (Sodunke et al 2007), spheroids (AbuAbsi et al 2002;Mueller et al 2011a), micro scaffolds (Bokhari et al 2007) or various types of hollow fiber bioreactors (Schmitmeier et al 2006;Schmelzer et al 2010). Hollow fiber 3D bioreactors were applied as bioartificial liver for clinical applications (Gerlach et al 2003) and were down-scaled for analytical purposes.…”

Section: Introductionmentioning

confidence: 99%

Real-time in situ viability assessment in a 3D bioreactor with liver cells using resazurin assay

et al. 2012

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Three-dimensional cultivation of human cells is promising especially for long-term maintenance of specific functions and mimicking the in vivo tissue environment. However, direct viability assessment is very difficult in such systems. Commonly applied indirect methods such as glucose consumption, albumin or urea production are greatly affected by culture conditions, stress and time of cultivation and do not reflect the real time viability of the cells. In this study we established a real-time in situ viability assay namely; resazurin assay, in a 3D hollow-fiber bioreactor using human liver cells. Resazurin assay is based on the conversion of resazurin to a fluorescent dye by cytoplasmatic and mitochondrial enzymes. We show that the resazurin reagent in concentrations used in this study is non-toxic and could be rapidly removed out of the system. Moreover, we observed that dead cells do not affect the results of the assay. We optimized the assay on HepG2 cells and tested it with primary human hepatocytes. Moreover, we maintained primary human hepatocytes in the 3D bioreactor system in serum-free conditions and also assessed viability before and after the exposure to amiodarone using the resazurin assay. We show that this approach is applicable during longterm cultivation of cells in bioreactors under different conditions and can moreover be applied to pharmacological studies, e.g. investigation of chronic drug effects in such 3D bioreactors.

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Micropatterns of Matrigel for three-dimensional epithelial cultures

Cited by 90 publications

References 31 publications

In vitro 3D angiogenesis assay in egg white matrix: comparison to Matrigel, compatibility to various species, and suitability for drug testing

In vitro 3D angiogenesis assay in egg white matrix: comparison to Matrigel, compatibility to various species, and suitability for drug testing

Three-dimensional cell culture: the missing link in drug discovery

Real-time in situ viability assessment in a 3D bioreactor with liver cells using resazurin assay

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