In Vitro High-Content Imaging-Based Phenotypic Analysis of Bronchial 3D Organotypic Air–Liquid Interface Cultures

Marescotti, Diego; Bovard, David; Morelli, Moran; Sandoz, Antonin; Luettich, Karsta; Frentzel, Stefan; Peitsch, Manuel C.; Hoeng, Julia

doi:10.1177/2472630319895473

Cited by 5 publications

(4 citation statements)

References 20 publications

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“…Furthermore, molecular target based drug screens are classically performed in 2D due to their ease of rapid assessment both through 2D image analysis and the use of high throughput high content multimode plate readers. However, these high throughput assays are difficult to translate to 3D, as confocal imaging is the standard imaging technique for cells in 3D yet is highly time intensive 13 . Thus, there is a need for 3D tissue engineered tumor microenvironments that can interface with high throughput drug screening methods and can assess anticancer drug efficacy in a rapid high content manner.…”

Section: Introductionmentioning

confidence: 99%

“…However, these high throughput assays are difficult to translate to 3D, as confocal imaging is the standard imaging technique for cells in 3D yet is highly time intensive. 13 Thus, there is a need for 3D tissue engineered tumor microenvironments that can interface with high throughput drug screening methods and can assess anticancer drug efficacy in a rapid high content manner. To bridge this gap, there has been a resurgence in phenotypic drug discovery.…”

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confidence: 99%

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Engineering high throughput screening platforms of cervical cancer

Cadena

Buchanan

Harris

et al. 2023

J Biomedical Materials Res

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Cervical cancer is the second leading cause of cancer‐related death in women under 40 and is one of the few cancers to have an increased incidence rate and decreased survival rate over the last 10 years. One in five patients will have recurrent and/or distant metastatic disease and these patients face a 5‐year survival rate of less than 17%. Thus, there is a pressing need to develop new anticancer therapeutics for this underserved patient population. However, the development of new anticancer drugs remains a challenge, as only 7% of novel anticancer drugs are approved for clinical use. To facilitate identification of novel and effective anticancer drugs for cervical cancer, we developed a multilayer multicellular platform of human cervical cancer cell lines and primary human microvascular endothelial cells that interfaces with high throughput drug screening methods to evaluate the anti‐metastatic and anti‐angiogenic drug efficacy simultaneously. Through the use of design of experiments statistical optimization, we identified the specific concentrations of collagen I, fibrinogen, fibronectin, GelMA, and PEGDA in each hydrogel layer that maximized both cervical cancer invasion and endothelial microvessel length. We then validated the optimized platform and assessed its viscoelastic properties. Finally, using this optimized platform, we conducted a targeted drug screen of four clinically relevant drugs on two cervical cancer cell lines. Overall, this work provides a valuable platform that can be used to screen large compound libraries for mechanistic studies, drug discovery, and precision oncology for cervical cancer patients.

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Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

Engineering high throughput screening platforms of cervical cancer

Cadena

Buchanan

Harris

et al. 2023

J Biomedical Materials Res

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“…S1) were acquired, respectively. Images were acquired with the CellInsight™ CX7 high-content screening platform (Thermo Fisher Scientific), and positively stained cells were quantified as previously described ( Marescotti et al, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Comparison of the basic morphology and function of 3D lung epithelial cultures derived from several donors

Bovard

Giralt

Trivedi

et al. 2020

Current Research in Toxicology

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In vitro models of the human lung play an essential role in evaluating the toxicity of inhaled compounds and understanding the development of respiratory diseases. Three-dimensional (3D) organotypic models derived from lung basal epithelial cells and grown at the air–liquid interface resemble human airway epithelium in multiple aspects, including morphology, cell composition, transcriptional profile, and xenobiotic metabolism. Whether the different characteristics of basal cell donors have an impact on model characteristics and responses remains unknown. In addition, studies are often conducted with 3D cultures from one donor, assuming a representative response on the population level. Whether this assumption is correct requires further investigation. In this study, we compared the morphology and functionality of 3D organotypic bronchial and small airway cultures from different donors at different weeks after air-lift to assess the interdonor variability in these parameters. The thickness, cell type composition, and transepithelial electrical resistance varied among the donors and over time after air-lift. Cilia beating frequency increased in response to isoproterenol treatment in both culture types, independent of the donor. The cultures presented low basal cytochrome P450 (CYP) 1A1/1B1 activity, but 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) treatment induced CYP1A1/1B1 activity regardless of the donor. In conclusion, lung epithelial cultures prepared from different donors present diverse morphology but similar functionality and metabolic activity, with certain variability in their response to stimulation.

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“…However, these high throughput assays are difficult to translate to 3D, as confocal imaging is the standard imaging technique for cells in 3D yet is highly time intensive. 18 Thus, there is a need for 3D tissue engineered tumor microenvironments that can interface with high throughput drug screening methods and can assess anti-cancer drug efficacy in a rapid high content manner. To bridge this gap, there has been a resurgence in phenotypic drug discovery.…”

Section: Introductionmentioning

confidence: 99%

Engineering High Throughput Screening Platforms of Cervical Cancer

Cadena

Buchanan

Harris

et al. 2022

Preprint

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There is a critical need for complex multicellular three-dimensional physiomimetic models of cancer that can interface with high throughput drug screening methods to assess anti-metastatic and anti-angiogenic drug efficacy in a rapid yet high content manner. We report a multilayer multicellular platform of human cervical cancer cell lines and primary human microvascular endothelial cells that incorporates critical biophysical and extracellular matrix cues, interfaces with standard high throughput drug screening methods, and can evaluate cervical cancer invasion and endothelial microvessel formation over time. Through the use of Design of Experiments statistical optimization, we identified the specific concentrations of collagen I, fibrinogen, fibronectin, GelMA, and PEGDA in each hydrogel layer that maximized cervical cancer invasion and endothelial microvessel length simultaneously. We then validated the optimized platform and assessed the viscoelastic properties of the composite hydrogels as well as their individual constituents. Finally, using this optimized platform, we conducted a targeted drug screen of four clinically relevant drugs on two cervical cancer cell lines. From these data we identified each of the cervical cancer cell lines (SiHa and Ca Ski) as either responsive or refractive to Paclitaxel, Dasitinib, Dovitinib, or Pazopanib. Overall, we developed a phenotypic drug screening platform of cervical cancer that captures cell behavior present in the cervical cancer tumor microenvironment, captures patient to patient variability, and integrates with standard high throughput high content drug screening methods. This work provides a valuable platform that can be used to screen large compound libraries for mechanistic studies, drug discovery, and precision oncology for cervical cancer patients.

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In Vitro High-Content Imaging-Based Phenotypic Analysis of Bronchial 3D Organotypic Air–Liquid Interface Cultures

Cited by 5 publications

References 20 publications

Engineering high throughput screening platforms of cervical cancer

Engineering high throughput screening platforms of cervical cancer

Comparison of the basic morphology and function of 3D lung epithelial cultures derived from several donors

Engineering High Throughput Screening Platforms of Cervical Cancer

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