Self-organization of human embryonic stem cells on micropatterns

Deglincerti, Alessia; Etoc, Fred; Martyn, Iain; Metzger, Jakob; Ruzo, Albert; Šimunović, Mijo; Yoney, Anna; Brivanlou, Ali H.; Siggia, Eric D.; Warmflash, Aryeh

doi:10.1038/nprot.2016.131

Cited by 137 publications

(153 citation statements)

References 31 publications

Supporting

Mentioning

147

Contrasting

Unclassified

Order By: Relevance

“…The lack of biophysical control of these preliminary models did not permit reproducibility or consistency required for experimental analysis. We sought to take advantage of cell micropatterning because of its’ proven ability to regulate stem cell fate and direct embryonic spatial patterning of pluripotent stem cells to model embryogenesis 9,10 . The current protocol of a robust long-term cell patterning method supplies biophysical confinement throughout the duration of the cardiac differentiation procedure to generate beating 3D cardiac microchambers to model the early developing human heart.…”

Section: Introductionmentioning

confidence: 99%

Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

et al. 2018

View full text Add to dashboard Cite

The discovery of human induced pluripotent stem cells (hiPSCs) has provided an unprecedented opportunity to study tissue morphogenesis and organ development as “organogenesis-in-a-dish”. Current approaches of cardiac organoid engineering rely on either direct cardiac differentiation from embryoid bodies or generation of aligned cardiac tissues from pre-differentiated cardiomyocytes from monolayer hiPSCs. To experimentally model early cardiac organogenesis in vitro, our protocol combines biomaterials-based cell patterning with stem cell organoid engineering. Three-dimensional cardiac microchambers are created from two-dimensional hiPSC colonies, which approximates an early developing heart with distinct spatial organization and self-assembly. With proper training on photolithography microfabrication, maintenance of human pluripotent stem cells, and cardiac differentiation, a graduate student with guidance will likely be able to carry out this experimental protocol, which requires approximately three weeks. We envisage that this in vitro model of human early heart development could serve as an embryotoxicity screening assay in drug discovery, regulation, and prescription for healthy fetal development. Currently, this protocol has yet to be implemented to assess the cardiac microchamber formation from the hiPSC lines with inherited disease, which can potentially be used to study the disease mechanisms of cardiac malformations at early stage of embryogenesis.

show abstract

Section: Introductionmentioning

confidence: 99%

Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Importantly, EMT plays a role in germ layer formation as epithelial epiblast cells ingress at the primitive streak (Stern, 2004). (Deglincerti et al, 2016b;Warmflash et al, 2014). Colors in patterned cell colonies represent different germ layers.…”

Section: The Role Of Adhesive Forcesmentioning

confidence: 99%

“…TE, trophectoderm. Microscopy images adapted with permission (Deglincerti et al, 2016b). and contraction, provide the mechanical force for complete embryo inversion (Hohn et al, 2015).…”

Section: Input From External Mechanical Cuesmentioning

confidence: 99%

Embryoids, organoids and gastruloids: new approaches to understanding embryogenesis

2017

Self Cite

View full text Add to dashboard Cite

Cells have an intrinsic ability to self-assemble and self-organize into complex and functional tissues and organs. By taking advantage of this ability, embryoids, organoids and gastruloids have recently been generated in vitro, providing a unique opportunity to explore complex embryological events in a detailed and highly quantitative manner. Here, we examine how such approaches are being used to answer fundamental questions in embryology, such as how cells selforganize and assemble, how the embryo breaks symmetry, and what controls timing and size in development. We also highlight how further improvements to these exciting technologies, based on the development of quantitative platforms to precisely follow and measure subcellular and molecular events, are paving the way for a more complete understanding of the complex events that help build the human embryo.

show abstract

“…39,40 Such bioengineered tissue samples may provide ideal templates for studying and modeling selforganized processes as biophysical models, such as limbs, organs, hair, and teeth. 76 Such studies would likely lead to advancements in regenerative medicine.…”

Section: Discussionmentioning

confidence: 99%

“…As a remark, similar edge effects were also found recently using human ESCs in vitro. 39,40 Such bioengineered tissue samples may provide ideal templates for studying or modeling a self-organized process for biophysical models, 2,41,42 contributing to the advancement of regenerative medicine.…”

mentioning

confidence: 99%

Role of the boundary in feather bud formation on one-dimensional bioengineered skin

Ishida

Mitsui

2018

APL Bioengineering

View full text Add to dashboard Cite

The role of a boundary in pattern formation from a homogenous state in Turing's reaction–diffusion equations is important, particularly when the domain size is comparable to the pattern scale. Such experimental conditions may be achieved for in vitro regeneration of ectodermal appendages such as feathers, via reconstruction of embryonic single cells. This procedure can eliminate a predefined genetic map, such as the midline of chick feather bud formation, leaving uniformly distributed identical cells as a bioengineered skin. Here, the self-organizing nature of multiple feather bud formation was examined in bioengineered 1D-skin samples. Primal formation of feather buds occurred at a fixed length from the skin edge. This formation was numerically recapitulated by a standard two-component reaction-diffusion model, suggesting that the boundary effect caused this observation. The proper boundary conditions were nonstandard, either mixed Dirichlet–Neumann or partial-flux. In addition, the model implies imperfect or hindered bud formation as well as nearly equal distances between buds. In contrast, experimental observations indicated that the skin curvature, which was not included in our model, also strongly affected bud formation. Thus, bioengineered skin may provide an ideal template for modeling a self-organized process from a homogenous state. This study will examine the possible diffusion activities of activator or inhibitor molecular candidates and mechanical activities during cell aggregation, which will advance our understanding of skin appendage regeneration from pluripotent or embryonic stem cells.

show abstract

Self-organization of human embryonic stem cells on micropatterns

Cited by 137 publications

References 31 publications

Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

Embryoids, organoids and gastruloids: new approaches to understanding embryogenesis

Role of the boundary in feather bud formation on one-dimensional bioengineered skin

Contact Info

Product

Resources

About