Revolutionizing Drug Discovery: Automating Patient-Derived Organoid (PDO) High-Content Assays with the BioAssemblyBot® (BAB) Platform

Hannah A. Strobel, PhD | Staff Scientist | Advanced Solutions Life Sciences 

Victoria Marsh, PhD | Director, Custom Organoid Services | Molecular Devices 

Maria R. Oyaga, PhD | Scientist | Core Life Analytics 

Angeline Lim, PhD | Applications Scientist | Molecular Devices 

Introduction: 

Patient-derived organoids (PDOs) are transforming how we approach drug and therapy development, offering more predictive models of patient responses. However, effectively utilizing these complex 3D tissue models for drug screening requires optimized workflows. Advanced Solutions Life Sciences, in collaboration with Molecular Devices and Core Life Analytics, has developed an innovative automation solution that significantly enhances PDO-based assays.    

The Challenge: Optimizing PDO-Based Drug Screening 

Traditional methods of handling and analyzing PDOs can be laborious, time-consuming, and prone to variability. This can hinder the throughput and reproducibility of drug screening efforts. There's a critical need for automation solutions that can streamline PDO workflows, improve data quality, and accelerate the drug discovery process.    

An Integrated BioAssemblyBot Solution:  

Advanced Solutions Life Sciences' BioAssemblyBot® platform offers a state-of-the-art automation solution designed to address these challenges. This agile system integrates several key components:    

• BioAssemblyBot® (BAB): A robotic system that automates the precise dispensing of PDOs and other liquid handling tasks.    

• BioStorageBot®: A modular incubator for automated cell culture.    

• BioApps™ Software: Intuitive workflow control software that integrates and manages all components of the platform.    

• Confocal Imaging System: For automated image acquisition of PDOs.    

• IN Carta® Image Analysis Software (Molecular Devices): For detailed image analysis.    

• StratoMineR™ (Core Life Analytics): For multiparametric analysis of the resulting data.    

Key Benefits of the BioAssemblyBot® Platform for PDO Assays: 

• Improved Accuracy and Reproducibility: The BAB's precise dispensing capabilities, including tunable parameters like pipetting speed, minimize organoid fragmentation and ensure uniform plating, leading to more consistent results.    

• Increased Throughput and Efficiency: Automation of key steps like PDO dispensing, media changes, and drug treatment significantly reduces processing time compared to manual methods.    

• Enhanced Data Quality: The integration of automated imaging and high-content analysis provides rich, multi-parametric data for a more comprehensive understanding of drug responses.    

• Workflow Simplification: The BioApps™ software provides a user-friendly interface to control and manage the entire automated workflow, making it easier to implement complex PDO assays.    

• Versatility: The platform is adaptable to a variety of PDO types and experimental designs.    

Case Study: Automated PDO Drug Screen for Colorectal Cancer 

The application note details a study where the BioAssemblyBot® platform was used to automate a drug screen of colorectal cancer PDOs.  

The workflow utilizing BAB included -  

1. Dispensing Matrigel domes containing PDOs 

2. Incubating PDOs in the BioStorageBot.    

3. Adding drug treatment (2 drugs with 4 or 5 dilutions) 

4. Aspirate culture medium and run staining protocol 

5. Automated confocal imaging.    

6. High-content image analysis and ATP production measurement.    

   
   
   
   

   

   Workflow for automated PDO drug screen.

   
   

   

   CRC PDOs seeded in Matrigel domes in a 96 well plate (A). PDOs after 7 days of culture (B) phase contrast image, and (C) calcein AM.

   
   

   

   TP production of PDOs, as measured by CellTiter-Glo 3D, performed at the end of the assay.

   
   

   

   Area of PDOs treated with different concentrations of trametinib or adavosertib, compared to untreated controls. Plates were seeded with PDOs from one of two different donors. Bars are mean ± SD.

   
   

   

   Heat maps and principal component analysis (PCA). For Donor 1 (PDO 1), 2 components are shown, which show limited clustering within or between drug concentrations. Donor 2, however, had clear clustering in components 1, 2, and 3, within each drug compared to controls.

   

   Box plots showing the Euclidian distance from the median of the control (untreated) for PDO 1 and PDO 2. Everything above the red dotted line has a p value <0.05.

The results demonstrated the BAB’s ability to: 

• Improve the accuracy and speed of Matrigel dome formation compared to manual pipetting.    

• Discriminate differences in drug response between PDOs from different patients.    

• Generate comprehensive data sets for in-depth analysis of drug effects.    

Conclusion 

The BioAssemblyBot® platform represents a significant advancement in automating PDO-based assays. By streamlining workflows, improving data quality, and increasing throughput, this technology empowers researchers to accelerate drug discovery and develop more personalized therapies.    

Acknowledgements:

Hannah A. Strobel, PhD | Staff Scientist | Advanced Solutions Life Sciences 

Victoria Marsh, PhD | Director, Custom Organoid Services | Molecular Devices 

Maria R. Oyaga, PhD | Scientist | Core Life Analytics 

Angeline Lim, PhD | Applications Scientist | Molecular Devices