An in vitro neurogenetics platform for precision disease modeling in the mouse DOI Creative Commons
Daniel Cortés, Mélanie Escudero, Austin C. Korgan

et al.

Science Advances, Journal Year: 2024, Volume and Issue: 10(14)

Published: April 3, 2024

The power and scope of disease modeling can be markedly enhanced through the incorporation broad genetic diversity. introduction pathogenic mutations into a single inbred mouse strain sometimes fails to mimic human disease. We describe cross-species precision platform that exploits diversity bridge cell-based with whole organism analysis. developed universal protocol permitted robust reproducible neural differentiation genetically diverse pluripotent stem cell lines then carried out proof-of-concept study neurodevelopmental gene DYRK1A . Results in vitro reliably predicted effects background on Dyrk1a loss-of-function phenotypes vivo. Transcriptomic comparison responsive unresponsive strains identified molecular pathways conferring sensitivity or resilience Dyrk1a1A loss highlighted differential messenger RNA isoform usage as an important determinant response. This strategy provides powerful tool functional analysis candidate variants studies.

Language: Английский

Advances in Human Cellular Mechanistic Understanding and Drug Discovery of Brain Organoids for Neurodegenerative Diseases DOI
Lei Tong, Xiaoshuang Zhang,

Gaoshuang Fu

et al.

Ageing Research Reviews, Journal Year: 2024, Volume and Issue: 102, P. 102517 - 102517

Published: Sept. 24, 2024

Language: Английский

Citations

3
Neuronal lineage tracing from progenitors in human cortical organoids reveals mechanisms of neuronal production, diversity, and disease DOI Creative Commons
Luke A. D. Bury, Shuai Fu, Anthony Wynshaw‐Boris

et al.

Cell Reports, Journal Year: 2024, Volume and Issue: 43(10), P. 114862 - 114862

Published: Oct. 1, 2024

Language: Английский

Citations

3
A Hybrid 2D‐to‐3D in vitro Differentiation Platform Improves Outcomes of Cerebral Cortical Organoid Generation in hiPSCs DOI
Dosh Whye,

Erika M. Norabuena,

Gayathri Srinivasan

et al.

Current Protocols, Journal Year: 2024, Volume and Issue: 4(10)

Published: Oct. 1, 2024

Abstract Three‐dimensional (3D) cerebral cortical organoids are popular in vitro cellular model systems widely used to study human brain development and disease, compared traditional stem cell–derived methods that use two‐dimensional (2D) monolayer cultures. Despite the advancements made protocol for organoid derivation over past decade, limitations due biological, mechanistic, technical variables remain generating these complex 3D systems. Building from our previously established differentiation system, we have modifications existing resolve several of biological challenges when working with diverse groups induced pluripotent cell (hiPSC) lines. This improved blends a 2D culture format specification neural cells expansion neuroepithelial progenitor system self‐aggregation subsequent development. Furthermore, this “hybrid” approach is amenable both an accelerated as well alternative long‐term protocol. In addition establishing hybrid format, also offers phenotypic morphological characterization stage‐specific profiles using antibodies fluorescent‐based dyes live imaging. © 2024 Wiley Periodicals LLC. Basic Protocol 1 : hiPSC‐based into (NSCs) 2 Serial passaging (NPCs) Support Direct cryopreservation rapid thawing NSCs NPCs 3 Bulk aggregation neurospheres Alternate High‐throughput neurosphere formation migration assay LIVE/DEAD stain imaging 4 NeuroFluor NeuO dye

Language: Английский

Citations

2
Cell-type-resolved somatic mosaicism reveals clonal dynamics of the human forebrain DOI Creative Commons
Changuk Chung, Xiaoxu Yang, Robert F. Hevner

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 25, 2023

Debate remains around anatomic origins of specific brain cell subtypes and lineage relationships within the human forebrain. Thus, direct observation in mature is critical for a complete understanding structural organization cellular origins. Here, we utilize mosaic variation types as distinct indicators clonal dynamics, denoted cell-type-specific Mosaic Variant Barcode Analysis. From four hemispheres from two different neurotypical donors, identified 287 780 variants (MVs), respectively that were used to deconvolve dynamics. Clonal spread allelic fractions reveal local hippocampal excitatory neurons are more lineage-restricted compared with resident neocortical or basal ganglia GABAergic inhibitory neurons. Furthermore, simultaneous genome-transcriptome analysis at both single-cell level suggests dorsal origin subgroup DLX1

Language: Английский

Citations

4
An in vitro neurogenetics platform for precision disease modeling in the mouse DOI Creative Commons
Daniel Cortés, Mélanie Escudero, Austin C. Korgan

et al.

Science Advances, Journal Year: 2024, Volume and Issue: 10(14)

Published: April 3, 2024

The power and scope of disease modeling can be markedly enhanced through the incorporation broad genetic diversity. introduction pathogenic mutations into a single inbred mouse strain sometimes fails to mimic human disease. We describe cross-species precision platform that exploits diversity bridge cell-based with whole organism analysis. developed universal protocol permitted robust reproducible neural differentiation genetically diverse pluripotent stem cell lines then carried out proof-of-concept study neurodevelopmental gene DYRK1A . Results in vitro reliably predicted effects background on Dyrk1a loss-of-function phenotypes vivo. Transcriptomic comparison responsive unresponsive strains identified molecular pathways conferring sensitivity or resilience Dyrk1a1A loss highlighted differential messenger RNA isoform usage as an important determinant response. This strategy provides powerful tool functional analysis candidate variants studies.

Language: Английский

Citations

1