Investigating the basis of lineage decisions and developmental trajectories in the dorsal spinal cord through pseudotime analyses DOI Creative Commons
Sandeep Gupta, Eric Heinrichs, Bennett G. Novitch

и другие.

Development, Год журнала: 2024, Номер 151(10)

Опубликована: Май 15, 2024

ABSTRACT Dorsal interneurons (dIs) in the spinal cord encode perception of touch, pain, heat, itchiness and proprioception. Previous studies using genetic strategies animal models have revealed important insights into dI development, but molecular details how dIs arise as distinct populations neurons remain incomplete. We developed a resource to investigate fate specification by combining single-cell RNA-Seq atlas mouse embryonic stem cell-derived with pseudotime analyses. To validate this silico useful tool, we used it first identify genes that are candidates for directing transition states lead lineage trajectories, then validated them situ hybridization analyses developing vivo. also identified an endpoint dI5 trajectory found become more transcriptionally homogeneous during terminal differentiation. This study introduces valuable tool further discovery about timing gene expression differentiation demonstrates its utility clarifying relationships.

Язык: Английский

Matrigel inhibits elongation and drives endoderm differentiation in aggregates of mouse embryonic stem cells DOI Creative Commons
Atoosa Amel, R. Michael Brown, Alexa Rabeling

и другие.

FEBS Open Bio, Год журнала: 2025, Номер unknown

Опубликована: Апрель 19, 2025

Modelling peri‐implantation mammalian development using the self‐organising properties of stem cells is a rapidly growing field that has advanced our understanding cell fate decisions occurring in early embryo. Matrigel, basement membrane matrix, critical substrate used various protocols for its efficacy promoting growth and self‐organisation. However, role driving lineage commitment, whether this effect driven by biochemical or physical cues, not currently clear. Here, we grow embryoid bodies suspension, Matrigel agarose, an inert polysaccharide, to attempt decouple roles better understand how it drives differentiation. We use combination light microscopy, quantitative PCR immunostaining investigate gene protein changes different culture conditions. show aggregates are hindered their ability elongate compared with those grown agarose indicating prohibitive Aggregates also differentiate into endoderm, ectoderm differentiation inhibited. Furthermore, these effects due presence as same witnessed agarose. Our results thus indicate significant complex on morphology bodies.

Язык: Английский

Процитировано

0
Signaling Centers Drive Brachyury Dynamics and Lineage Commitment in mESC Aggregates DOI Creative Commons
Alexa Rabeling, Mubeen Goolam

FASEB BioAdvances, Год журнала: 2025, Номер unknown

Опубликована: Май 2, 2025

ABSTRACT Stem cell‐based models of embryogenesis have exploded in popularity, resulting protocols with overlapping use some reagents and differential others. As such, the precise contributions individual signaling molecules, such as Chiron BMP4, applied to whole or part aggregates, matrices, Matrigel, development these are unknown. Furthermore, different methods, matrices has yet be directly compared under same conditions. In this paper, we used a mouse embryonic stem cell aggregate model compare BMP4 pulses (applied aggregate) centers embedded them low‐percentage Matrigel. Each factor had effects on morphology, Brachyury protein expression, lineage commitment, having pulses. pulse was shown drive neural differentiation, while resulted better recapitulation aspects anterior–posterior axis formation, polarization expression anterior posterior genes observed. This further elucidates inform decisions around experimental conditions for vitro development.

Язык: Английский

Процитировано

0
In preprints: exploring developmental robustness and timing with gastruloids DOI
Yonit Maroudas-Sacks, Marc Trani Bustos, Jesse V. Veenvliet

и другие.

Development, Год журнала: 2025, Номер 152(10)

Опубликована: Май 15, 2025

Язык: Английский

Процитировано

0
A modular organization of mammalian gastrulation and the Spemann-Mangold organizer DOI
Alfonso Martínez Arias, André Dias, Alexandra E. Wehmeyer

и другие.

Cells and Development, Год журнала: 2025, Номер unknown, С. 204031 - 204031

Опубликована: Май 1, 2025

Язык: Английский

Процитировано

0
Chimeric 3D gastruloids – a versatile tool for studies of mammalian peri-gastrulation development DOI Open Access
Alexandra E. Wehmeyer, Katrin M. Schüle, Alexandra Conrad

и другие.

Development, Год журнала: 2022, Номер 149(22)

Опубликована: Ноя. 3, 2022

ABSTRACT Stem cell-derived three-dimensional (3D) gastruloids show a remarkable capacity of self-organisation and recapitulate many aspects gastrulation stage mammalian development. Gastruloids can be rapidly generated offer several experimental advantages, such as scalability, observability accessibility for manipulation. Here, we present approaches to further expand the potency murine 3D by using functional genetics in mouse embryonic stem cells (mESCs) generate chimeric gastruloids. In gastruloids, fluorescently labelled different genotypes harbouring inducible gene expression or loss-of-function alleles are combined with wild-type cells. We showcase this approach mESCs carrying homozygous deletions Tbx transcription factor brachyury Eomes. Resulting reported Eomes functions, instructing cardiac fate promoting posterior axial extension, respectively. Additionally, revealed previously unrecognised phenotypes, tissue sorting preference deficient endoderm cell non-autonomous effects deficiency on Wnt3a patterning along axis, demonstrating some advantages an efficient tool studies gastrulation.

Язык: Английский

Процитировано

15
Negative feedback on Retinoic Acid by Brachyury guides gastruloid symmetry-breaking DOI Creative Commons

Meagan J. Hennessy,

Timothy Fulton, David A. Turner

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Июнь 4, 2023

Abstract Establishment of the vertebrate body plan requires a combination extra-embryonic signalling to establish morphogen gradients, and an underlying self-assembly mechanism that contributes pattern regulation robustness. Gastruloids are aggregates mouse embryonic stem cells break morphological symmetry polarise Brachyury ( Bra ) expression in absence signals. However, by which breaking occurs is not yet known. During gastrulation axis elongation, retinoic acid (RA) Cyp26a1 polarised along anteroposterior axis, this critical for balancing decision self-renew or differentiate. We found symmetry-breaking gastruloids coincident with separation Aldh1a2 expression, feedback from maintaining gene gastruloid posterior region. Furthermore, we reveal short temporal window where RA can negatively influence both expression. These observations lead us suggest how initial patterning, subsequent evolving network topologies create defined boundaries permits proper axial patterning growth.

Язык: Английский

Процитировано

8
Integrated Molecular-Phenotypic Profiling Reveals Metabolic Control of Morphological Variation in Stembryos DOI Creative Commons
Alba Villaronga Luque, Ryan Savill, Natalia López-Anguita

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Дек. 5, 2023

SUMMARY Mammalian stem-cell-based models of embryo development (stembryos) hold great promise in basic and applied research. However, considerable phenotypic variation despite identical culture conditions limits their potential. The biological processes underlying this seemingly stochastic are poorly understood. Here, we investigate the roots by intersecting transcriptomic states morphological history individual stembryos across stages modeling post-implantation early organogenesis. Through machine learning integration time-resolved single-cell RNA-sequencing with imaging-based quantitative profiling, identify features predictive end-state. Leveraging power revealed that imbalance oxidative phosphorylation glycolysis results aberrant morphology a neural lineage bias can be corrected metabolic interventions. Collectively, our work establishes divergent as drivers variation, offers broadly applicable framework to chart predict organoid systems. strategy leveraged control processes, ultimately increasing reproducibility vitro Highlights Time-resolved charting hundreds generates molecular fingerprints Machine identifies end-state Early cellular composition Metabolic interventions tune stembryo correct derailment differentiation outcomes

Язык: Английский

Процитировано

8
From signalling oscillations to somite formation DOI Creative Commons
Wilke H. M. Meijer, Katharina F. Sonnen

Current Opinion in Systems Biology, Год журнала: 2024, Номер 39, С. 100520 - 100520

Опубликована: Май 25, 2024

Periodic segmentation of vertebrate embryos or somitogenesis is regulated by a dynamic network signalling pathways. Signalling gradients determine the spacing forming segments, while oscillations, collectively termed clock, ensure their regular timing. Since clock paradigm dynamics at tissue level, its mechanism and function have been topic many studies. Recently, researchers able to analyse quantify these with unprecedented precision, revealing complexity interlinked oscillations tissue-wide throughout development. Initial studies shown how interplay between cellular mechanics drive periodic formation segments. Looking ahead, new techniques such as in vitro stem cell-based models (human) embryonic development will enable detailed investigations into mechanisms somitogenesis.

Язык: Английский

Процитировано

2
The logic of monsters: development and morphological diversity in stem-cell-based embryo models DOI Creative Commons
Dominica Cao, Sumit Garai, James DiFrisco

и другие.

Interface Focus, Год журнала: 2024, Номер 14(5)

Опубликована: Окт. 25, 2024

Organoids and stem-cell-based embryo models (SEMs) are imperfect organ or representations that explore a much larger space of possible forms, morphospace, compared to their

Язык: Английский

Процитировано

2
OptoRheo: Simultaneous in situ micro-mechanical sensing and imaging of live 3D biological systems DOI Creative Commons
Tania Mendonca, Katarzyna Lis-Slimak, Andrew B. Matheson

и другие.

Communications Biology, Год журнала: 2023, Номер 6(1)

Опубликована: Апрель 28, 2023

Abstract Biomechanical cues from the extracellular matrix (ECM) are essential for directing many cellular processes, normal development and repair, to disease progression. To better understand cell-matrix interactions, we have developed a new instrument named ‘OptoRheo’ that combines light sheet fluorescence microscopy with particle tracking microrheology. OptoRheo lets us image cells in 3D as they proliferate over several days while simultaneously sensing mechanical properties of surrounding pericellular at sub-cellular length scale. can be used two operational modalities (with without an optical trap) extend dynamic range microrheology measurements. We corroborated this by characterising ECM live breast cancer distinct culture systems, cell clusters hydrogels spheroids suspension culture. This cutting-edge will transform exploration drug transport through complex matrices optimise design next-generation models.

Язык: Английский

Процитировано

5