The differentiation of human induced pluripotent stem cells (hiPSCs) into specific cell types for disease modeling and restorative therapies is a key research agenda and offers the possibility to obtain patient-specific cells of interest for a wide range of diseases. Basal forebrain cholinergic neurons (BFCNs) play a particular role in the pathophysiology of Alzheimer's dementia and isolated dystonias. In this work, various directed differentiation protocols based on monolayer neural induction were tested for their effectiveness in promoting a ventral telencephalic phenotype and generating BFCN. Ventralizing factors [i.e., purmorphamine and Sonic hedgehog (SHH)] were applied at different time points, time intervals, and concentrations. In addition, caudal identity was prevented by the use of a small molecule XAV-939 that inhibits the Wnt-pathway. After patterning, gene expression profiles were analyzed by quantitative PCR (qPCR). Rostro-ventral patterning is most effective when initiated simultaneously with neural induction. The most promising combination of patterning factors was 0.5 μM of purmorphamine and 1 μM of XAV-939, which induces the highest expression of transcription factors specific for the medial ganglionic eminence, the source of GABAergic inter- and cholinergic neurons in the telencephalon. Upon maturation of cells, the immune phenotype, as well as electrophysiological properties were investigated showing the presence of marker proteins specific for BFCN (choline acetyltransferase, ISL1, p75, and NKX2.1) and GABAergic neurons. Moreover, a considerable fraction of measured cells displayed mature electrophysiological properties. Synaptic boutons containing the vesicular acetylcholine transporter (VACHT) could be observed in the vicinity of the cells. This work will help to generate basal forebrain interneurons from hiPSCs, providing a promising platform for modeling neurological diseases, such as Alzheimer's disease or Dystonia.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8416478 | PMC |
| http://dx.doi.org/10.3389/fcell.2021.716249 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
INTERSPECIES ORGANOIDS REVEAL HUMAN-SPECIFIC MOLECULAR FEATURES OF DOPAMINERGIC NEURON DEVELOPMENT AND VULNERABILITY.
bioRxiv
November 2024
The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA.
The disproportionate expansion of telencephalic structures during human evolution involved tradeoffs that imposed greater connectivity and metabolic demands on midbrain dopaminergic neurons. Despite the central role of dopaminergic neurons in human-enriched disorders, molecular specializations associated with human-specific features and vulnerabilities of the dopaminergic system remain unexplored. Here, we establish a phylogeny-in-a-dish approach to examine gene regulatory evolution by differentiating pools of human, chimpanzee, orangutan, and macaque pluripotent stem cells into ventral midbrain organoids capable of forming long-range projections, spontaneous activity, and dopamine release.
View Article and Find Full Text PDFFGF8-mediated gene regulation affects regional identity in human cerebral organoids.
Elife
November 2024
Univ. Côte d'Azur (UniCA), CNRS, Inserm, Institut de Biologie Valrose (iBV), Nice, France.
Article Synopsis
- The study investigates how the morphogen FGF8 affects the development of brain-like structures derived from human stem cells, specifically focusing on its role in establishing cellular diversity and regional identities during early brain development.
- FGF8 treatment led to the formation of various brain regions within the organoids, influencing the identity of neural progenitor cells and the ratio of different types of neurons (GABAergic and glutamatergic).
- The research highlights FGF8's critical role in regulating genes linked to neurodevelopmental disorders, suggesting its potential involvement in both healthy brain development and related pathological conditions.
Thalamus of Reptiles and Mammals: Some Significant Differences.
Brain Behav Evol
October 2024
Department of Biomedical Engineering, University of Utah, Salt Lake City, Utah, USA.
Background: Most studies comparing forebrain organization between reptiles and mammals have focused on similarities. Equally important are the differences between their brains. While differences have been addressed infrequently, this approach can highlight the evolution of brains in relation to their respective environments.
View Article and Find Full Text PDFAstrocyte allocation during brain development is controlled by Tcf4-mediated fate restriction.
EMBO J
November 2024
Department of Anesthesia, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, and Zhongshan Hospital, Fudan University, Shanghai, 200032, China.
Article Synopsis
- Astrocytes in the brain are distributed unequally across regions, affecting advanced brain functions, but how they are allocated is not fully understood.
- This study reveals that the transcription factor 4 (Tcf4) plays a critical role in determining where astrocytes are positioned during development, based on their origin from embryonic cells.
- Loss of Tcf4 leads to misplacement of astrocytes in the wrong brain regions and may contribute to issues seen in neurodevelopmental disorders associated with Tcf4 mutations.
Huntington's disease cellular phenotypes are rescued non-cell autonomously by healthy cells in mosaic telencephalic organoids.
Nat Commun
August 2024
Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122, Milan, Italy.
Article Synopsis
- Huntington's disease (HD) leads to the degeneration of specific brain neurons, resulting in a mix of functional and dysfunctional cells, but the interactions between these cell types are not well understood.
- Researchers created brain organoids containing both healthy and HD cells, finding that HD organoids displayed neurodevelopmental issues and fewer GABAergic neurons compared to healthy ones.
- Healthy cells in mixed organoids helped to restore the identity and function of HD cells through direct interactions, suggesting that enhancing communication between different cell types could offer new treatment strategies for HD.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!