Acoustic Droplet-Assisted Superhydrophilic–Superhydrophobic Microarray Platform for High-Throughput Screening of Patient-Derived Tumor Spheroids

2021 ◽  
Author(s):  
Yu Xia ◽  
Hui Chen ◽  
Juan Li ◽  
Hang Hu ◽  
Qun Qian ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microarray Platform ◽  
Tumor Spheroids

scholarly journals Nano-biofilm Arrays as a Novel Universal Platform for Microscale Microbial Culture and High-Throughput Downstream Applications

2019 ◽  
Vol 26 (14) ◽  
pp. 2529-2535 ◽  
Author(s):  
Anand Srinivasan ◽  
Anand K. Ramasubramanian ◽  
José L. Lopez-Ribot
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Antimicrobial Agents ◽  
Microarray Platform ◽  
New Drugs ◽  
Microbial Culture ◽  
Phenotypic Properties ◽  
Microbial Biofilms ◽  
Antibiotic Drug

Biofilms are the predominant mode of microbial growth and it is now fully accepted that a majority of infections in humans are associated with a biofilm etiology. Biofilms are defined as attached and structured microbial communities surrounded by a protective exopolymeric matrix. Importantly, sessile microorganisms growing within a biofilm are highly resistant to antimicrobial agents. Thus, there is an urgent need to develop new and improved anti-biofilm therapies. Unfortunately, most of the current techniques for in-vitro biofilm formation are not compatible with high throughput screening techniques that can speed up discovery of new drugs with anti-biofilm activity. To try to overcome this major impediment, our group has developed a novel technique consisting of micro-scale culture of microbial biofilms on a microarray platform. Using this technique, hundreds to thousands of microbial biofilms, each with a volume of approximately 30-50 nanolitres, can be simultaneously formed on a standard microscope slide. Despite more than three orders of magnitude of miniaturization over conventional biofilms, these nanobiofilms display similar growth, structural and phenotypic properties, including antibiotic drug resistance. These nanobiofilm chips are amenable to automation, drastically reducing assay volume and costs. This technique platform allows for true high-throughput screening in search for new anti-biofilm drugs.

High Throughput Screening of Cell Mechanical Response Using a Stretchable 3D Cellular Microarray Platform

Small ◽  
2020 ◽  
Vol 16 (30) ◽  
pp. 2000941 ◽  
Author(s):  
Kabilan Sakthivel ◽  
Hitendra Kumar ◽  
Mohamed G. A. Mohamed ◽  
Bahram Talebjedi ◽  
Justin Shim ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Mechanical Response ◽  
Microarray Platform

scholarly journals A sandwiched microarray platform for benchtop cell-based high throughput screening

Biomaterials ◽  
2011 ◽  
Vol 32 (3) ◽  
pp. 841-848 ◽  
Author(s):  
Jinhui Wu ◽  
Ian Wheeldon ◽  
Yuqi Guo ◽  
Tingli Lu ◽  
Yanan Du ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microarray Platform

scholarly journals Evaluation of the Droplet-Microarray Platform for High-Throughput Screening of Suspension Cells

2016 ◽  
Vol 22 (2) ◽  
pp. 163-175 ◽  
Author(s):  
Anna A. Popova ◽  
Claire Depew ◽  
Katya Manuella Permana ◽  
Alexander Trubitsyn ◽  
Ravindra Peravali ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microarray Platform ◽  
Free Cell ◽  
Suspension Cells ◽  
Cell Content ◽  
Microtiter Plates ◽  
Microarray Screening ◽  
Screening Platform ◽  
Rare Cells

Phenotypic cell-based high-throughput screenings play a central role in drug discovery and toxicology. The main tendency in cell screenings is the increase of the throughput and decrease of reaction volume in order to accelerate the experiments, reduce the costs, and enable screenings of rare cells. Conventionally, cell-based assays are performed in microtiter plates, which exist in 96- to 1536-wells formats and cannot be further miniaturized. In addition, performing screenings of suspension cells is associated with risk of losing cell content during the staining procedures and incompatibility with high-content microscopy. Here, we evaluate the Droplet-Microarray screening platform for culturing, screening, and imaging of suspension cells. We demonstrate pipetting-free cell seeding and proliferation of cells in individual droplets of 3–80 nL in volume. We developed a methodology to perform parallel treatment, staining, and fixation of suspension cells in individual droplets. Automated imaging of live suspension cells directly in the droplets combined with algorithms for pattern recognition for image analysis is demonstrated. We evaluated the developed methodology by performing a dose–response study with antineoplastic drugs. We believe that the DMA screening platform carries great potential to be adopted for broad spectrum of screenings of suspension cells.

scholarly journals RNAi High-Throughput Screening of Single- and Multi-Cell-Type Tumor Spheroids: A Comprehensive Analysis in Two and Three Dimensions

2017 ◽  
Vol 22 (5) ◽  
pp. 525-536 ◽  
Author(s):  
Jiaqi Fu ◽  
Daniel Fernandez ◽  
Marc Ferrer ◽  
Steven A. Titus ◽  
Eugen Buehler ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Attrition Rate ◽  
Three Dimensions ◽  
Cell Type ◽  
Tumor Spheroids ◽  
Flat Bottom ◽  
Cancer Subtypes

The widespread use of two-dimensional (2D) monolayer cultures for high-throughput screening (HTS) to identify targets in drug discovery has led to attrition in the number of drug targets being validated. Solid tumors are complex, aberrantly growing microenvironments that harness structural components from stroma, nutrients fed through vasculature, and immunosuppressive factors. Increasing evidence of stromally-derived signaling broadens the complexity of our understanding of the tumor microenvironment while stressing the importance of developing better models that reflect these interactions. Three-dimensional (3D) models may be more sensitive to certain gene-silencing events than 2D models because of their components of hypoxia, nutrient gradients, and increased dependence on cell-cell interactions and therefore are more representative of in vivo interactions. Colorectal cancer (CRC) and breast cancer (BC) models composed of epithelial cells only, deemed single-cell-type tumor spheroids (SCTS) and multi-cell-type tumor spheroids (MCTS), containing fibroblasts were developed for RNAi HTS in 384-well microplates with flat-bottom wells for 2D screening and round-bottom, ultra-low-attachment wells for 3D screening. We describe the development of a high-throughput assay platform that can assess physiologically relevant phenotypic differences between screening 2D versus 3D SCTS, 3D SCTS, and MCTS in the context of different cancer subtypes. This assay platform represents a paradigm shift in how we approach drug discovery that can reduce the attrition rate of drugs that enter the clinic.

scholarly journals 3D hanging spheroid plate for high-throughput CAR T cell cytotoxicity assay

2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Zhenzhong Chen ◽  
Seokgyu Han ◽  
Arleen Sanny ◽  
Dorothy Leung-Kwan Chan ◽  
Danny van Noort ◽  
...  
Keyword(s):  
T Cells ◽  
Tumor Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Tumor Type ◽  
Tumor Spheroids ◽  
Cytotoxic Effects ◽  
Cytotoxicity Assays ◽  
Car T Cells ◽  
Car T

Abstract Background Most high-throughput screening (HTS) systems studying the cytotoxic effect of chimeric antigen receptor (CAR) T cells on tumor cells rely on two-dimensional cell culture that does not recapitulate the tumor microenvironment (TME). Tumor spheroids, however, can recapitulate the TME and have been used for cytotoxicity assays of CAR T cells. But a major obstacle to the use of tumor spheroids for cytotoxicity assays is the difficulty in separating unbound CAR T and dead tumor cells from spheroids. Here, we present a three-dimensional hanging spheroid plate (3DHSP), which facilitates the formation of spheroids and the separation of unbound and dead cells from spheroids during cytotoxicity assays. Results The 3DHSP is a 24-well plate, with each well composed of a hanging dripper, spheroid wells, and waste wells. In the dripper, a tumor spheroid was formed and mixed with CAR T cells. In the 3DHSP, droplets containing the spheroids were deposited into the spheroid separation well, where unbound and dead T and tumor cells were separated from the spheroid through a gap into the waste well by tilting the 3DHSP by more than 20°. Human epidermal growth factor receptor 2 (HER2)-positive tumor cells (BT474 and SKOV3) formed spheroids of approximately 300–350 μm in diameter after 2 days in the 3DHSP. The cytotoxic effects of T cells engineered to express CAR recognizing HER2 (HER2-CAR T cells) on these spheroids were directly measured by optical imaging, without the use of live/dead fluorescent staining of the cells. Our results suggest that the 3DHSP could be incorporated into a HTS system to screen for CARs that enable T cells to kill spheroids formed from a specific tumor type with high efficacy or for spheroids consisting of tumor types that can be killed efficiently by T cells bearing a specific CAR. Conclusions The results suggest that the 3DHSP could be incorporated into a HTS system for the cytotoxic effects of CAR T cells on tumor spheroids. Graphical Abstract

scholarly journals 3D Hanging Spheroid Plate for High-Throughput CAR T Cell Cytotoxicity Assay

2021 ◽  
Author(s):  
Zhenzhong Chen ◽  
Seokgyu Han ◽  
Arleen Sanny ◽  
Dorothy Leung-Kwan Chan ◽  
Danny van Noort ◽  
...  
Keyword(s):  
T Cells ◽  
Tumor Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Tumor Type ◽  
Tumor Spheroids ◽  
Cytotoxic Effects ◽  
Cytotoxicity Assays ◽  
Car T Cells ◽  
Car T

Abstract Background Most high-throughput screening (HTS) systems for the cytotoxic effect of chimeric antigen receptor (CAR) T cells on tumor cells rely on two-dimensional cell culture that does not suitably recapitulate the tumor microenvironment (TME). Tumor spheroids can recapitulate TME and have been used for cytotoxicity assays of CAR T cells. However, a major obstacle to the use of tumor spheroids for cytotoxicity assays is the difficulty in separating unbound CAR T and dead tumor cells from spheroids. Here, we present a three-dimensional hanging spheroid plate (3DHSP), which facilitates the formation of spheroids and the separation of unbound and dead cells from spheroids during cytotoxicity assays. Results The 3DHSP is a 24-well plate, with each well composed of a hanging dripper, spheroid wells, and waste wells. In the dripper, a tumor spheroid was formed and mixed with CAR T cells. In 3DHSP, droplets containing the spheroids were deposited into the spheroid separation well, where unbound and dead T and tumor cells were separated from the spheroid through a gap into the waste well by tilting the 3DHSP by more than 20°. Human epidermal growth factor receptor 2 (HER2)-positive tumor cells (BT474 and SKOV3) formed spheroids of approximately 300–350 mm in diameter after 2 d in the 3DHSP. The cytotoxic effects of T cells engineered to express CAR recognizing HER2 (HER2-CAR T cells) on these spheroids were directly measured by optical imaging, without the use of live and dead fluorescent staining of the cells. Our results suggest that the 3DHSP could be incorporated into an HTS system to screen for CARs that enable T cells to kill spheroids formed from a specific tumor type with high efficacy or for spheroids consisting of tumor types that can be killed efficiently by T cells bearing a specific CAR. Conclusions The results suggest that the 3DHSP could be incorporated into an HTS system for the cytotoxic effects of CAR T cells on tumor spheroids.

scholarly journals A Novel Multiparametric Drug-Scoring Method for High-Throughput Screening of 3D Multicellular Tumor Spheroids Using the Celigo Image Cytometer

2017 ◽  
Vol 22 (5) ◽  
pp. 547-557 ◽  
Author(s):  
Scott Cribbes ◽  
Sarah Kessel ◽  
Scott McMenemy ◽  
Jean Qiu ◽  
Leo Li-Ying Chan
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Method ◽  
Three Dimensional ◽  
Cancer Drug ◽  
Multicellular Tumor Spheroids ◽  
Tumor Spheroids ◽  
Scoring Method ◽  
Drug Candidates ◽  
Drug Compounds

Three-dimensional (3D) tumor models have been increasingly used to investigate and characterize cancer drug compounds. The ability to perform high-throughput screening of 3D multicellular tumor spheroids (MCTS) can highly improve the efficiency and cost-effectiveness of discovering potential cancer drug candidates. Previously, the Celigo Image Cytometer has demonstrated a novel method for high-throughput screening of 3D multicellular tumor spheroids. In this work, we employed the Celigo Image Cytometer to examine the effects of 14 cancer drug compounds on 3D MCTS of the glioblastoma cell line U87MG in 384-well plates. Using parameters such as MCTS diameter and invasion area, growth and invasion were monitored for 9 and 3 d, respectively. Furthermore, fluorescent staining with calcein AM, propidium iodide, Hoechst 33342, and caspase 3/7 was performed at day 9 posttreatment to measure viability and apoptosis. Using the kinetic and endpoint data generated, we created a novel multiparametric drug-scoring system for 3D MCTS that can be used to identify and classify potential drug candidates earlier in the drug discovery process. Furthermore, the combination of quantitative and qualitative image data can be used to delineate differences between drugs that induce cytotoxic and cytostatic effects. The 3D MCTS-based multiparametric scoring method described here can provide an alternative screening method to better qualify tested drug compounds.

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