Cell Mechanics in Embryoid Bodies

Zeevaert, Kira; Mabrouk, Mohamed H Elsafi; Wagner, Wolfgang; Goetzke, Roman

doi:10.3390/cells9102270

Cited by 45 publications

(66 citation statements)

References 95 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Notably, at early stages of cell aggregates the self-detached EBs revealed higher self-organization than Spin-EBs, which might be due to their generation from pre-organized iPSC colonies. Albeit the subsequent growth and multilineage differentiation of self-detached EBs and Spin-EBs was very similar, it is conceivable, that there are functional differences that warrant further analysis between different methods for EB formation (Pettinato et al, 2015; Zeevaert et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

The spatial self-organization within pluripotent stem cell colonies is continued in detaching aggregates

Mabrouk

Goetzke

Abagnale

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Colonies of induced pluripotent stem cells (iPSCs) reveal aspects of self-organization even under culture conditions that maintain pluripotency. To investigate the dynamics of this process under spatial confinement, we used either polydimethylsiloxane (PDMS) pillars or micro-contact printing of vitronectin. There was a progressive upregulation of OCT4, E-cadherin, and NANOG within 70 micro meter from the outer rim of iPSC colonies. Single-cell RNA-sequencing demonstrated that OCT4high subsets have pronounced up-regulation of the TGF-beta pathway, particularly of NODAL and its inhibitor LEFTY, at the rim of the colonies. Furthermore, calcium-dependent cell-cell interactions were found to be relevant for the self-organization. Interestingly, after 5 to 7 days, the iPSC colonies detached spontaneously from micro-contact printed substrates to form 3D aggregates. This new method allowed generation of embryoid bodies (EBs) of controlled size, without any enzymatic or mechanical treatment. Within the early 3D aggregates, the radial organization and differential gene expression continued in analogy to the changes observed during self-organization of iPSC colonies. Our results provide further insight into the gradual self-organization within iPSC colonies and at their transition into EBs.

show abstract

Section: Discussionmentioning

confidence: 99%

The spatial self-organization within pluripotent stem cell colonies is continued in detaching aggregates

Mabrouk

Goetzke

Abagnale

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…During EB formation rather homogeneous epithelial-like hPSC colonies undergo epithelial-mesenchymal transition (EMT) and develop into 3D structures containing interacting cells indicative of the ectoderm, mesoderm and endoderm. Although in principal, EBs can be used for target identification and drug testing, one must be aware that they lack controlled organization of differentiation or tissue development and do neither recapitulate a realistic in vivo condition nor a specific stage or morphological structure of an embryo [ 20 , 53 , 54 , 55 , 56 ].…”

Section: Human Organoidsmentioning

confidence: 99%

Human Embryo Models and Drug Discovery

Rosner¹,

Reithofer²,

Fink³

et al. 2021

IJMS

View full text Add to dashboard Cite

For obvious reasons, such as, e.g., ethical concerns or sample accessibility, model systems are of highest importance to study the underlying molecular mechanisms of human maladies with the aim to develop innovative and effective therapeutic strategies. Since many years, animal models and highly proliferative transformed cell lines are successfully used for disease modelling, drug discovery, target validation, and preclinical testing. Still, species-specific differences regarding genetics and physiology and the limited suitability of immortalized cell lines to draw conclusions on normal human cells or specific cell types, are undeniable shortcomings. The progress in human pluripotent stem cell research now allows the growth of a virtually limitless supply of normal and DNA-edited human cells, which can be differentiated into various specific cell types. However, cells in the human body never fulfill their functions in mono-lineage isolation and diseases always develop in complex multicellular ecosystems. The recent advances in stem cell-based 3D organoid technologies allow a more accurate in vitro recapitulation of human pathologies. Embryoids are a specific type of such multicellular structures that do not only mimic a single organ or tissue, but the entire human conceptus or at least relevant components of it. Here we briefly describe the currently existing in vitro human embryo models and discuss their putative future relevance for disease modelling and drug discovery.

show abstract

“…Given the significant influence of spatial cues in directing human fetal development and hPSC differentiation, and prior work connecting embryoid body size with downstream spheroid and organoid development [ 22 ], we hypothesized that shape may also influence hCO development and brain region specification and that these changes would be reflected in differential gene expression patterns. To test this, we engineered agarose microwells with distinct shapes but equivalent volumes and seeded embryoid bodies (EB) within them.…”

Section: Introductionmentioning

confidence: 99%

Effects of early geometric confinement on the transcriptomic profile of human cerebral organoids

2021

View full text Add to dashboard Cite

Background Human cerebral organoids (hCO) are attractive systems due to their ability to model important brain regions and transcriptomics of early in vivo brain development. To date, they have been used to understand the effects of genetics and soluble factors on neurodevelopment. Interestingly, one of the main advantages of hCOs are that they provide three dimensionality that better mimics the in vivo environment; yet, despite this central feature it remains unclear how spatial and mechanical properties regulate hCO and neurodevelopment. While biophysical factors such as shape and mechanical forces are known to play crucial roles in stem cell differentiation, embryogenesis and neurodevelopment, much of this work investigated two dimensional systems or relied on correlative observations of native developing tissues in three dimensions. Using hCOs to establish links between spatial factors and neurodevelopment will require the use of new approaches and could reveal fundamental principles of brain organogenesis as well as improve hCOs as an experimental model. Results Here, we investigated the effects of early geometric confinements on transcriptomic changes during hCO differentiation. Using a custom and tunable agarose microwell platform we generated embryoid bodies (EB) of diverse shapes mimicking several structures from embryogenesis and neurodevelopment and then further differentiated those EBs to whole brain hCOs. Our results showed that the microwells did not have negative gross impacts on the ability of the hCOs to differentiate towards neural fates, and there were clear shape dependent effects on neural lineage specification. In particular we observed that non-spherical shapes showed signs of altered neurodevelopmental kinetics and favored the development of medial ganglionic eminence-associated brain regions and cell types over cortical regions. Transcriptomic analysis suggests these mechanotransducive effects may be mediated by integrin and Wnt signaling. Conclusions The findings presented here suggest a role for spatial factors in brain region specification during hCO development. Understanding these spatial patterning factors will not only improve understanding of in vivo development and differentiation, but also provide important handles with which to advance and improve control over human model systems for in vitro applications.

show abstract

Cell Mechanics in Embryoid Bodies

Cited by 45 publications

References 95 publications

The spatial self-organization within pluripotent stem cell colonies is continued in detaching aggregates

The spatial self-organization within pluripotent stem cell colonies is continued in detaching aggregates

Human Embryo Models and Drug Discovery

Effects of early geometric confinement on the transcriptomic profile of human cerebral organoids

Contact Info

Product

Resources

About