organoid cell cultures

How automated organoid cell cultures are developed, imaged, and analyzed

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3D cell models are becoming increasingly popular for studying complex biological effects, tissue functionality, and diseases. Their ability to self-organize and mimic functional organ cell types is believed to better represent in vivo biology than 2D monolayer cell cultures. While the complexity of these models can be a hurdle for wider use in research and drug development, barriers are being broken down with innovative technologies.

In our latest post, scientists at Molecular Devices discuss advances in cell technologies and demonstrate an integrated workflow that allows automated processes of cell culture and imaging to monitor the development and characterize the complex responses in 3D organoids.

High-content imaging methods for automated cell culture organoids

When using organoids for disease modeling and assessment of compound effects, the quality of images is important for downstream analysis. For maximum quantitative and robust assessment of phenotypic changes in organoids, as well as for increasing throughput of experiments and screens, high-performance, automated imaging and analysis solutions are of critical importance.

Confocal microscopy enables efficient imaging of 3D objects, including spheroids, organoids, and organ-on-a-chip models. A confocal microscope, such as the ImageXpress® Confocal HT.ai High-Content Imaging System utilizes a seven-channel laser light source with eight imaging channels to enable highly multiplexed assays; spinning disc confocal technology to penetrate deeper into thick tissue samples for sharper images; and water immersion objective increase signal to noise ratio, improve z-resolution, and decrease optical aberrations for sharper, crisper images

Traditional image analysis methods can be incredibly intricate and time-consuming when performed manually or even semi-automatically. There’s always the possibility of human error and bias due to the task’s complex and highly detailed nature. When you add to this the repetitive, lengthy, and often laborious nature of the workflow, there comes the opportunity to apply advanced image analysis tools and AI/machine learning.

Advanced image analysis software would provide information about changes in phenotypes. MetaXpress® High-Content Image and Acquisition and Analysis Software allows users to find and characterize spheroids and then count and characterize cells inside spheroids, as well as sub-cellular objects. You can assess the specifics of 3D cell cultures, such as volume, diameter, shape, and intensity, which also helps you classify and organize data according to organoid features.

IN Carta® Image Analysis Software is a deep learning-based image segmentation tool enables robust label-free organoids and cell analysis. The machine learning tools can convert complex image data into actionable results. This solution helped researchers classify organoids based on size and diameter.

Overall, our proprietary AgileOptix technologies in confocal microscopy boast the qualities that can elaborately map out the complex structure of 3D organoids.

Organoid applications for drug discovery and development

Organoids are becoming increasingly important in the fields of cancer research, neurobiology, stem cell research, and drug discovery, since they allow for the enhanced modeling of human tissues. Derived from stem cells, organoids can be differentiated into a wide range of tissue types including the lung, brain, and intestine to name a few. Because these 3D microtissues mimic in vivo organs, they can provide researchers with greater insight into the mechanisms of human development and disease, for example:

Pulmonary (lung) organoids

Lung organoid cultures are 3D microtissue models recapitulating the morphological and functional characteristics of the airway, which include alveolar structure, mucus secretion, ciliary beating, and regeneration. These special characteristics of the lung organoid culture hold potential for a wide range of applications in both basic and translational approaches such as drug screening and disease modeling.

Pulmonary (lung) organoids

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Intestinal (gut) organoids

Intestinal organoids or gut organoids were one of the first 3D organoid models to recapitulate structures in the intestinal lumen and on the surrounding intestinal epithelium. The cell composition and arrangement of the epithelium make intestinal organoids useful for studying intestinal cell biology, regeneration, differentiation, as wells as disease phenotypes including effects of specific mutations, microbiome, or inflammation process.

Intestinal (gut) organoids

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Brain (cerebral) organoids

Brain organoids are 3D tissue models representing one or more regions of the brain. When cultured, human-induced pluripotent stem cells (hiPSCs) differentiate into various neural cells that mature over time to resemble structures of various brain regions, Cerebral 3D organoids are a rapidly developing technology that has great potential for understanding human brain development, neuronal diseases, and can be used for testing the effects of compounds and genetic mutations. This approach is highly promising for the evaluation of pharmaceutical drugs, studying effects of toxins, and in functional genomic applications.

Brain (cerebral) organoids

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PDO/Tumoroids

Patient-derived organoids (PDOs) – or tumoroids – are 3D cultures that can be generated from primary tumors of individual patients. Tumoroids are highly valuable tools for cancer research, drug development, and personalized medicine.

For example, efficient cancer therapy is crucial in the survival of cancer patients. This necessitates the use of clinically-relevant tumor models to understand the biology of disease, analyze tumor biomarkers, screen for the most efficient anti-cancer drugs, and provide a platform to study responses to targeted therapies.

PDO/Tumoroids

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Automation protocol of 3D biology workflows

Due to the complexity of organoids, more sophisticated 3D imaging and analysis techniques are needed to characterize these biological assays accurately and efficiently. Today, automated confocal imaging systems and 3D image analysis software are commonly used to help researchers streamline their workflow and obtain optimal results.

Our organoid screening workflow demonstrates an end-to-end method that utilizes leading technology from across the industry for automated cell culture, monitoring, and high-content imaging. The integrated workcell includes our SpectraMax® microplate reader, Aquamax microplate washer, ImageXpress® Confocal high-content imagers, automated CO2 incubator, automated liquid handler, as well as a collaborative robot. With intuitive scheduling software, researchers can control the 3D workflow for automating the seeding, media exchange, and monitoring of organoids development. In addition, the method allows for automation of compound testing and evaluation of phenotypic changes.

Automation protocol of 3D biology workflows
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At our Organoid Innovation Center (OIC) we showcase these cutting-edge technologies with novel 3D biology methods to address key challenges of scaling complex 3D biology. The collaborative space brings customers and researchers into the lab to test automated workflows for organoid culturing and screening, with guidance from in-house scientists.

Taking a closer look at automated organoid development

To learn more about specific applications of organoids and the importance of lab automation in organoid workflows, join our latest webinar, Automating Culture and High-Content Imaging of 3D Organoids for in Vitro Assessment of Compound Effects. The webinar includes case studies with different organoid types used in tissue modeling and drug screening. You will gain valuable information about how automated cell culture organoids are developed, imaged, analyzed, and understand the role our imaging and software solutions play throughout organoid research.

Automating Culture and High-Content Imaging of 3D Organoids

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