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Use of 3D Organoids as a Model to Study Idiopathic Form of Parkinson's Disease.
Organoids are becoming particularly popular in modeling diseases that are difficult to reproduce in animals, due to anatomical differences in the structure of a given organ. Thus, they are a bridge between the in vitro and in vivo models. Human midbrain is one of the structures that is currently being intensively reproduced in organoids for modeling Parkinson's disease (PD). Thanks to three-dimensional (3D) architecture and the use of induced pluripotent stem cells (iPSCs) differentiation into organoids, it has been possible to recapitulate a complicated network of dopaminergic neurons. In this work, we present the first organoid model for an idiopathic form of PD. iPSCs were generated from peripheral blood mononuclear cells of healthy volunteers and patients with the idiopathic form of PD by transduction with Sendai viral vector. iPSCs were differentiated into a large multicellular organoid-like structure. The mature organoids displayed expression of neuronal early and late markers. Interestingly, we observed statistical differences in the expression levels of LIM homeobox transcription factor alpha (early) and tyrosine hydroxylase (late) markers between organoids from PD patient and healthy volunteer. The obtained results show immense potential for the application of 3D human organoids in studying the neurodegenerative disease and modeling cellular interactions within the human brain.
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A scalable 3D packaging technique for brain organoid arrays toward high-capacity bioprocessors.
Neural organoids provide a promising platform for biologically inspired computing due to their complex neural architecture and energy-efficient signal processing. However, the scalability of conventional organoid cultures is limited, restricting synaptic connectivity and functional capacity-significant barriers to developing high-performance bioprocessors. Here, we present a scalable three-dimensional (3D) packaging strategy for neural organoid arrays inspired by semiconductor 3D stacking technology. This approach vertically assembles Matrigel-embedded neural organoids within a polydimethylsiloxane (PDMS)-based chamber using a removable acrylic alignment plate, creating a stable multilayer structure while preserving oxygen and nutrient diffusion. Structural analysis confirms robust inter-organoid connectivity, while electrophysiological recordings reveal significantly enhanced neural dynamics in 3D organoid arrays compared to both single organoids and two-dimensional arrays. Furthermore, prolonged culture duration promotes network maturation and increases functional complexity. This 3D stacking strategy provides a simple yet effective method for expanding the physical and functional capacity of organoid-based systems, offering a viable path toward next-generation biocomputing platforms.
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Single Cell Data Enables Dissecting Cell Types Present in Bulk Transcriptome Data.
The quality of organoid models can be assessed by single-cell-RNA-sequencing (scRNA-seq) but often only bulk transcriptome data is available. Here we present a pipeline for the analysis of scRNA-seq data and subsequent "deconvolution," which is a method for estimating cell type fractions in bulk transcriptome data based on expression profiles and cell types found in scRNA-seq data derived from biopsies. We applied this pipeline on bulk iPSC-derived kidney and brain organoid transcriptome data to identify cell types employing two scRNA-seq kidney datasets and one brain dataset. Relevant cells present in kidney (e.g., proximal tubules, distal convoluted tubules, and podocytes) and brain (e.g., neurons, astrocytes, oligodendrocytes, and microglia) with obligatory endothelial and immune-related cells were identified. We anticipate that this pipeline will also enable estimation of cell type fractions in organoids of other tissues.
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Developmental neuroscience: Miniature human brains.
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What Implications Do a Consciousness-Independent Perspective on Moral Status Entail for Future Brain Organoid Research?
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These 3D model brains with cells from several people are first of their kind.
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Persistent organic pollutants: Neurotoxic mechanisms and insights from organoid models.
Persistent Organic Pollutants (POPs) pose significant global environmental health risks due to their persistence, bioaccumulation, and long-range transport. Exposure to POPs has been linked to a range of neurological disorders, including cognitive impairment, neurodevelopmental abnormalities, and neurodegenerative diseases. This review examines the neurotoxic effects of various POP categories, such as flame retardants, pesticides, plasticizers, and preservatives, and explores the underlying molecular mechanisms. It also highlights the potential of brain organoids as a more physiologically relevant experimental platform. Although organoid models offer improved translational value, their capacity to fully replicate the complexity of the human nervous system remains limited, necessitating further technical refinement and interdisciplinary collaboration. The integration of organoid technology into environmental toxicology is anticipated to enhance the accuracy and efficiency of neurotoxicity assessment, thereby supporting the development of robust health risk evaluation frameworks and informing evidence-based environmental policy.
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Neurodevelopment. Lab dishes up mini-brains.
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Uniform cerebral organoid culture on a pillar plate by simple and reproducible spheroid transfer from an ultralow attachment well plate.
Human induced pluripotent stem cell (iPSC)-derived brain organoids have potential to recapitulate the earliest stages of brain development, serving as an effectivein vitromodel for studying both normal brain development and disorders. However, current brain organoid culture methods face several challenges, including low throughput, high variability in organoid generation, and time-consuming, multiple transfer and encapsulation of cells in hydrogels throughout the culture. These limitations hinder the widespread application of brain organoids including high-throughput assessment of compounds in clinical and industrial lab settings. In this study, we demonstrate a straightforward approach of generating multiple cerebral organoids from iPSCs on a pillar plate platform, eliminating the need for labor-intensive, multiple transfer and encapsulation steps to ensure the reproducible generation of cerebral organoids. We formed embryoid bodies in an ultra-low attachment 384-well plate and subsequently transferred them to the pillar plate containing Matrigel, using a straightforward sandwiching and inverting method. Each pillar on the pillar plate contains a single spheroid, and the success rate of spheroid transfer was in a range of 95%-100%. Using this approach, we robustly generated cerebral organoids on the pillar plate and demonstrated an intra-batch coefficient of variation below 9%-19% based on ATP-based cell viability and compound treatment. Notably, our spheroid transfer method in combination with the pillar plate allows miniaturized culture of cerebral organoids, alleviates the issue of organoid variability, and has potential to significantly enhance assay throughput by allowingin situorganoid assessment as compared to conventional organoid culture in 6-/24-well plates, petri dishes, and spinner flasks.
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Lineage Recording in Human Brain Organoids with iTracer.
Induced pluripotent stem cell (iPSC)-derived organoids provide models to study human organ development. Single-cell transcriptomics enables highly resolved descriptions of cell states within these systems; however, approaches are needed to directly determine the lineage relationship between cells. Here we provide a detailed protocol (Fig. 1) for the application of iTracer (He Z, Maynard A, Jain A, et al., Nat Methods 19:90-99, 2022), a recently published lineage recorder that combines reporter barcodes with inducible CRISPR-Cas9 scarring and is compatible with single-cell and spatial transcriptomics. iTracer is used to explore clonality and lineage dynamics during brain organoid development. More broadly, iTracer can be adapted to any iPSC-derived culture system to dissect lineage dynamics during normal or perturbed development.