Media additives to promote spheroid circularity and compactness in hanging drop platform

2015 ◽  
Vol 3 (2) ◽  
pp. 336-344 ◽  
Author(s):  
Brendan M. Leung ◽  
Sasha Cai Lesher-Perez ◽  
Toshiki Matsuoka ◽  
Christopher Moraes ◽  
Shuichi Takayama
Keyword(s):  
High Throughput ◽  
Drug Screening ◽  
Three Dimensional ◽  
Hanging Drop

Three-dimensional spheroid cultures have become increasingly popular as drug screening platforms, especially with the advent of different high throughput spheroid forming technologies.

Patient-derived organoids for personalized drug screening in intrahepatic cholangiocarcinoma.

2020 ◽  
Vol 38 (4_suppl) ◽  
pp. 581-581
Author(s):  
Ricardo J. Antonia ◽  
Kan Toriguchi ◽  
Eveliina Karelehto ◽  
Dania Annuar ◽  
Luika Timmerman ◽  
...  
Keyword(s):  
High Throughput ◽  
Drug Screening ◽  
Intrahepatic Cholangiocarcinoma ◽  
Three Dimensional ◽  
Small Samples ◽  
Patient Specific ◽  
Mtor Inhibition ◽  
Drug Responses

581 Background: Despite standard treatment with gemcitabine and cisplatin, median survival for unresectable Intrahepatic Cholangiocarcinoma (ICC) is < 1 year. Clearly, novel therapeutic strategies are urgently needed. The paucity of targetable mutations in ICC and the as yet unproven benefit of genetically targeted drugs led us to ask whether a reliable clinical benefit may be revealed by patient-specific therapeutic testing in novel models of ICC. Here we describe our ability to establish patient-derived three-dimensional organoid cultures (PDO) that enable individualized identification of active single agents or drug combinations in surrogate models of ICC. Methods: To model patient-specific drug responses, we used the freshly resected ICCs from small samples of single patient tumors to generate PDXs and PDOs, small spheroidal clusters of tumor cells grown in vitro. We have employed a high-throughput drug screening platform using AI-enhanced robotics (Yamaha Motor Corporation) to identify and distribute single, uniformly sized PDOs into 384-well ultra-low adherent plates. This is coupled with a TECAN D300e drug dispenser that rapidly delivers nanoliter volumes of a 34-drug panel, thereby facilitating rapid, reliable drug response analyses. Results: Our data show that PDOs retain characteristic genomic and histological features of the patients’ tumors. Drug responses were specific to each patient tumor, but PDOs from all patients responded to a greater or lesser degree to mTOR inhibition, suggesting that this pathway is important in ICC. The responses of PDO to the mTOR inhibitor Sapanisertib (INK128), was recapitulated in the same patient’s PDX. Further, INK128 was synergistic with gemcitabine in patient 970 PDOs as well as in vivo in PDX also from patient 970. Conclusions: As it is believed that PDX can predict patient responses to drugs, our results suggest that PDO may also predict patient drug responses. The establishment of PDO may allow economical patient-specific, high throughput drug screens that could ultimately inform clinical practice. [Table: see text]

Abstract 997: Three-dimensional high-throughput drug screening for bladder cancer

2019 ◽  
Author(s):  
Sang-Cheol Lee ◽  
Young Saing Kim ◽  
In Gyu Hwang ◽  
Su Jin Lee ◽  
Se Hoon Park
Keyword(s):  
Bladder Cancer ◽  
High Throughput ◽  
Drug Screening ◽  
Three Dimensional ◽  
High Throughput Drug Screening

Abstract 997: Three-dimensional high-throughput drug screening for bladder cancer

2019 ◽  
Author(s):  
Sang-Cheol Lee ◽  
Young Saing Kim ◽  
In Gyu Hwang ◽  
Su Jin Lee ◽  
Se Hoon Park
Keyword(s):  
Bladder Cancer ◽  
High Throughput ◽  
Drug Screening ◽  
Three Dimensional ◽  
High Throughput Drug Screening

scholarly journals Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening

10.3791/57476 ◽  
2018 ◽  
Author(s):  
Lesley Mathews Griner ◽  
Kalyani Gampa ◽  
Toan Do ◽  
Huyen Nguyen ◽  
David Farley ◽  
...  
Keyword(s):  
High Throughput ◽  
Drug Screening ◽  
Three Dimensional ◽  
Tumor Spheroids

scholarly journals Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging

2021 ◽  
Vol 7 (17) ◽  
pp. eabc1323
Author(s):  
A. Ganguli ◽  
A. Mostafa ◽  
C. Saavedra ◽  
Y. Kim ◽  
P. Le ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Drug Screening ◽  
Three Dimensional ◽  
3D Culture ◽  
Geometric Control ◽  
Hanging Drop ◽  
Design Elements ◽  
Trade Offs ◽  
On Chip

Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from specific matrices. Microchip cultures from human LN229 glioblastoma and patient-derived mouse xenograft cells retained genomic alterations of originating tumors based on mate pair sequencing. We measured response to drugs over time with real-time microscopy on-chip. Last, by engineering droplets to form predetermined geometric shapes, we were able to manipulate the geometry of cultured cell masses. These outcomes can enable broad applications in advancing personalized medicine for cancer and drug discovery, tissue engineering, and stem cell research.

scholarly journals A Facile High-Throughput Model of Surface-Independent Staphylococcus aureus Biofilms by Spontaneous Aggregation

mSphere ◽  
2021 ◽  
Vol 6 (2) ◽  
Author(s):  
Terrence Cheng ◽  
Nelson S. Torres ◽  
Ping Chen ◽  
Anand Srinivasan ◽  
Sandra Cardona ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Staphylococcus Aureus ◽  
High Throughput ◽  
Three Dimensional ◽  
Hanging Drop ◽  
Methicillin Resistant ◽  
Content Type ◽  
Throughput Model

ABSTRACT Many microbes in their natural habitats are found in biofilm ecosystems attached to surfaces and not as free-floating (planktonic) organisms. Furthermore, it is estimated that nearly 80% of human infections are associated with biofilms. Biofilms are traditionally defined as three-dimensional, structured microbial communities that are attached to a surface and encased in a matrix of exopolymeric material. While this view of biofilm largely arises from in vitro studies under static or flow conditions, in vivo observations have indicated that this view of biofilms is essentially true only for foreign-body infections on catheters or implants where biofilms are attached to the biomaterial. In mucosal infections such as chronic wounds or cystic fibrosis or joint infections, biofilms can be found unattached to a surface and as three-dimensional aggregates. In this work, we describe a high-throughput model of aggregate biofilms of methicillin-resistant Staphylococcus aureus (MRSA) using 96-well plate hanging-drop technology. We show that MRSA forms surface-independent biofilms, distinct from surface-attached biofilms, that are rich in exopolymeric proteins, polysaccharides, and extracellular DNA (eDNA), express biofilm-related genes, and exhibit heightened antibiotic resistance. We also show that the surface-independent biofilms of clinical isolates of MRSA from cystic fibrosis and central catheter-related infections demonstrate morphological differences. Overall, our results show that biofilms can form by spontaneous aggregation without attachment to a surface, and this new in vitro system can model surface-independent biofilms that may more closely mimic the corresponding physiological niche during infection. IMPORTANCE The canonical model of biofilm formation begins with the attachment and growth of microbial cells on a surface. While these in vitro models reasonably mimic biofilms formed on foreign bodies such as catheters and implants, this is not the case for biofilms formed in cystic fibrosis and chronic wound infections, which appear to present as aggregates not attached to a surface. The hanging-drop model of biofilms of methicillin-resistant Staphylococcus aureus (MRSA), the major causative organism of skin and soft tissue infections, shows that these biofilms display morphological and antibiotic response patterns that are distinct from those of their surface-attached counterparts, and biofilm growth is consistent with their in vivo location. The simplicity and throughput of this model enable adoption to investigate other single or polymicrobial biofilms in a physiologically relevant setting.

scholarly journals Pulsed Laser Assisted High-Throughput Intracellular Delivery in Hanging Drop Based Three Dimensional Cancer Spheroids

The Analyst ◽  
2021 ◽  
Author(s):  
Pallavi Gupta ◽  
Srabani Kar ◽  
Ashish Kumar ◽  
Fan-Gang Tseng ◽  
Shantanu Pradhan ◽  
...  
Keyword(s):  
High Throughput ◽  
Three Dimensional ◽  
Pulsed Laser ◽  
Intracellular Delivery ◽  
Invasive Technique ◽  
Hanging Drop ◽  
Cellular Processes ◽  
Non Invasive ◽  
Cancer Spheroids

Targeted intracellular delivery of biomolecules and therapeutic cargo enables controlled manipulation of cellular processes. Laser-based optoporation has emerged as a versatile, non-invasive technique that employs light-based transient physical disruption of...

Organ-on-Chip Devices Toward Applications in Drug Development and Screening

2018 ◽  
Vol 12 (4) ◽  
Author(s):  
Christopher Uhl ◽  
Wentao Shi ◽  
Yaling Liu
Keyword(s):  
Drug Development ◽  
High Throughput ◽  
Drug Screening ◽  
Large Scale ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Throughput Capacity ◽  
Comprehensive Overview ◽  
The Past ◽  
High Throughput Drug Screening

As a necessary pathway to man-made organs, organ-on-chips (OOC), which simulate the activities, mechanics, and physiological responses of real organs, have attracted plenty of attention over the past decade. As the maturity of three-dimensional (3D) cell-culture models and microfluidics advances, the study of OOCs has made significant progress. This review article provides a comprehensive overview and classification of OOC microfluidics. Specifically, the review focuses on OOC systems capable of being used in preclinical drug screening and development. Additionally, the review highlights the strengths and weaknesses of each OOC system toward the goal of improved drug development and screening. The various OOC systems investigated throughout the review include, blood vessel, lung, liver, and tumor systems and the potential benefits, which each provides to the growing challenge of high-throughput drug screening. Published OOC systems have been reviewed over the past decade (2007–2018) with focus given mainly to more recent advances and improvements within each organ system. Each OOC system has been reviewed on how closely and realistically it is able to mimic its physiological counterpart, the degree of information provided by the system toward the ultimate goal of drug development and screening, how easily each system would be able to transition to large scale high-throughput drug screening, and what further improvements to each system would help to improve the functionality, realistic nature of the platform, and throughput capacity. Finally, a summary is provided of where the broad field of OOCs appears to be headed in the near future along with suggestions on where future efforts should be focused for optimized performance of OOC systems in general.

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