Brain Organoids
Brain organoids are 3D tissue models derived from human-induced pluripotent stem cells (hiPSCs). When cultured, the stem cells differentiate into various neural cells that mature over time to resemble structures of brain regions such as the forebrain (cerebral) or mid-brain.
The culture of complex organoids such as the cerebral organoid is a rapidly developing technology that has immense potential in areas of developmental neurobiology and neurodegeneration. Recent progress has made it possible to culture brain organoids from iPSCs which allows for the study of cortical development and the diseases associated with it. Further work is required before cerebral organoids can be used at scale for functional genomics studies, drug discovery, and the study of compound-induced toxicity effects.
General steps used to develop iPSC-derived cerebral organoids
Cerebral organoids were grown according to the protocol adapted from Lancaster & Knoblich, 2014. In this method, the cerebral organoids are grown in culture media that promotes self-organization and patterning.
- Day 0 – Embryoid Bodies (EB) formation - iPSC cells were seeded in a 96-well ultra-low attachment plate
- Day 2-5 – Germ layer differentiation - feeding and monitoring the EBs
- Day 5 – Neural induction - transfer to 24-well plate containing neural induction medium
- Day 7 – Transfer to Matrigel droplets - neuroectodermal tissues are transferred to droplets of Matrigel
- Day 8-10 – Neuroepithelial bud expansion - the outgrowth of neuroepithelial buds expand and contain fluid-filled lumens
- Day 11 – Maturation - tissues are transferred to a spinning bioreactor to promote growth and expansion
Monitor developing 3D cerebral organoids using AI-based analysis tools
Developing brain microtissues can be monitored for their size and morphology using AI-based analysis tools like our IN Carta® Image Analysis Software, helping to define the size and shape of the tissues. Selected microtissues were analyzed by confocal imaging during different phases of development for cellular organization by expression of different neuronal markers.
Figure 2: Analysis of brightfield images using deep-learning based segmentation. A) EBs were monitored for their sizes before proceeding to the neural induction step. Shown are examples of images and their corresponding segmentation masks using the SINAP tool in IN Carta software. B) Histogram shows the distribution of EB diameter. Binned size = 10 μm. C) Maturation of organoids can be monitored using brightfield imaging and analyzed using IN Carta SINAP tool. Scale bar = 100 μm.
Calcium imaging of brain organoids to determine neuronal activity
For detection of functional activities, organoids were loaded with calcium-sensitive dye, and then Ca2+ oscillations were recorded with the ImageXpress® Confocal HT.ai High-Content Imaging System and analyzed using MetaXpress® High- Content Image Acquisition and Analysis Software for kinetic peak analysis. We show that high-content imaging paired with AI-based analysis used with 3D cerebral organoids is a promising tool for compound screens and toxicity evaluations.
Figure 3. Synchronous calcium activity by week 13 . A) Image shows an optical section of calcium 6 loaded organoid. B) Calcium intensity from boxed region is represented as average intensity over time. C) Close-up view of box region from (A) shown over time. Arrows indicate elevated intensity. Bottom images represent calcium intensity as a heat map. Note the intensity spreading from the initial spike suggesting the presence of neuronal network.
Featured asset
Automated monitoring of development and activity analysis of iPSC-derived 3D cerebral organoids
Here, we describe a method for the semi-automated culture and monitoring of cerebral organoids, as well as testing functional neuronal activity by means of recording Ca2+ oscillations. Brightfield imaging with artificial intelligence (AI)-based segmentation can help monitor the quality of developing organoids by tracking the growth in diameter and shape. The neuronal activity of brain organoids can be determined from calcium activity. Confocal imaging reveals calcium activity to determine the maturity of the neurons. Furthermore, the cellular organization can be monitored via confocal imaging with differential staining.
Latest resources on organoid research
The complexity of 3D assays remains a hurdle for the wider adoption of organoid models in research and drug screening. New advanced tools for imaging and analysis, as well as assay automation are critical for increased quality of information, throughput, and precision of complex biological models. Learn how high-throughput, high-content imaging and analysis paired with AI-based analytic tools can improve the accuracy of your organoid studies.
Resources for Brain Organoids
Blog
Stem Cell Science and Regenerative Medicine – Technology and Methods Presented at ISSCR 2022
Stem Cell Science and Regenerative Medicine – Technology and Methods Presented at ISSCR 2022
It was another great year at ISSCR 2022! Leaders from across the globe came together to discuss new technologies, share insights, and explore the newest breakthroughs in stem cell…
Application Note
Neurite outgrowth analysis in 3D reconstructed human brain micro-tissues
Neurite outgrowth analysis in 3D reconstructed human brain micro-tissues
Organotypic 3D cultures resemble native 3D tissue architecture and are believed more representative of real tissues than their 2D (monolayer) counterparts, thus providing higher in vivo…
Scientific Poster
Automated monitoring of development and activity analysis of iPSC-derived 3D cerebral organoids
Automated monitoring of development and activity analysis of iPSC-derived 3D cerebral organoids
The culture of complex organoids such as the cerebral organoid is a rapidly developing technology that has immense potential in areas of developmental neurobiology and neurodegeneration.
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Blog
Engineering Next-gen Organoids with Automated Lab Workflows at #SLAS2022
Engineering Next-gen Organoids with Automated Lab Workflows at #SLAS2022
SLAS2022, the Society for Lab Automation and Screening conference offered another exciting year for learning about innovative laboratory technologies. Whether you attended in-person…
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Scientific Poster
Automation of 3D organoid culture workflow with deep-learning based image analysis
Automation of 3D organoid culture workflow with deep-learning based image analysis
3D cell culture as a model system is increasingly popular because it recapitulates the in vivo microenvironment better than 2D cell cultures. Organoids have the capacity for stable different…
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Blog
History of Organoid Research: From Sponge Cells to Functional Organs
History of Organoid Research: From Sponge Cells to Functional Organs
Let's begin with a simple definition of an organoid which refers to a three-dimensional assembly that contains multiple cell types that are arranged with realistic histology, at…
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Blog
Stem cell science insights and breakthroughs presented at #ISSCR2021
Stem cell science insights and breakthroughs presented at #ISSCR2021
If you didn't get a chance to visit us at our poster sessions during ISSCR 2021, don’t fret. We've gathered all our sessions right here for you. The ISSCR Annual Meeting brought…
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Scientific Poster
Monitoring organoid development and characterization of calcium oscillation activities in iPSC-derived 3D cerebral organoids
Monitoring organoid development and characterization of calcium oscillation activities in iPSC-derived 3D cerebral organoids
Cerebral organoids are a rapidly developing technology that has great potential for understanding brain development and neuronal diseases. They can also be used for testing effects of compou…
Publications
Project Profile: Organoid Innovation Center
Project Profile: Organoid Innovation Center
Lab Manager speaks to Dan O’Connor, vice president, drug discovery, Molecular Devices, about the company’s Organoid Innovation Center in San Jose, CA. The center is 180 sq. ft., within a lar…
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Application Note
Functional evaluation of neurotoxic and neuroactive compound effects in iPSC-derived 3D neural co-cultures
Functional evaluation of neurotoxic and neuroactive compound effects in iPSC-derived 3D neural co-cultures
There is increasing interest in using more complex, biologically-relevant, and predictive cell-based platforms for assay development and compound screening.
Customer Breakthrough
StemoniX uses three of our solutions to validate their microBrain 3D Assay Ready Plates
StemoniX uses three of our solutions to validate their microBrain 3D Assay Ready Plates
Current models available to neuroscience researchers have limitations that hamper the development of new medicines. Complex models, such as post-mortem brains and animal models, more closely…
Videos & Webinars
Application and Analysis of Organoid Systems
StemoniX microBrain 3D Assay Ready Plates for HTS
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