Directed Assembly and Development of Material‐Free Tissues with Complex Architectures

Vrij, E.J.; Rouwkema, Jeroen; LaPointe, Vanessa; Blitterswijk, Clemens van; Truckenmüller, Roman; Rivron, Nicolas

doi:10.1002/adma.201505723

Cited by 56 publications

(64 citation statements)

References 45 publications

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“…Microfabrication Elastomeric stamps for imprinting the agarose microwells were fabricated using PDMS Sylgard 184 kit. Microwells were molded using a 2.2% w/v solution of Ultrapure agarose (thermo fisher scientific 11560166) as described previously (Vrij et al, 2016a). Note for 3D cell cultures: each well of a 96-wellplate contains a total of 430 microwells with a calculated liquid volume of 250 µl split between 225 µl medium and 25 µl hydrogel buffer.…”

Section: Methodsmentioning

confidence: 99%

“…We used a high-content screening platform of non-adherent hydrogel microwells in 96 well-plates (Vrij et al, 2016a) to reproducibly aggregate small number of ESCs into EBs (Figure 1A). ESCs seeded into microwells were normally distributed across the 430 microwells within each well (7-12 cells per microwell) and formed EBs within 24 hours (Figure 1B, S1).…”

Section: Naïve Pluripotency Increase Pre Differentiationmentioning

confidence: 99%

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Chemically-defined induction of a primitive endoderm and epiblast-like niche supports post-implantation progression from blastoids

et al. 2019

Preprint

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The early mammalian embryo (blastocyst) contains three co-developing types of stem cells. Two supporting extraembryonic types -the trophectoderm and the primitive endoderm -encase and guide the pluripotent epiblast that eventually forms all body tissues. Unlike embryos, stem cell-based models of the embryo can be generated in large numbers and subjected to high-content screens as a basis for basic and biomedical discoveries (Rivron et al., 2018a;Vrij et al., 2016b). Here, we show that aggregates of naive Embryonic stem cells cultured in chemically-defined conditions and exposed to combinatorial screens of signaling molecules, rapidly (48 hours) and efficiently (80%) set apart PrE-like cells. These cells produce a basal lamina, generate progenitors resembling both visceral and parietal endoderm, and polarize co-developing Epiblast cells to form a proamniotic cavity. In blastoids, stem cell-based models of the early blastocyst (Rivron et al., 2018b), this combination of signals increases the ratio and number of Gata6+/Nanog+ cells and promotes the survival, growth and morphogenesis of a post-implantation-like Epiblast in vitro. Modeling early embryonic development in chemically-defined in vitro conditions shows that the primitive endoderm forms via a specific combination of signaling pathways and sufficient to drive the development of the Epiblast. Hamazaki et al., 2004). However, EBs are formed in chemically-undefined conditions and PrE is not generated efficiently, which occlude studying underlying mechanisms. Recently, we reported the generation of blastoids (Rivron et al., 2018b), embryonic stem cell (ESC) and trophoblast stem cell (TSC)-based structures resembling the E3.5 blastocyst. Building on previous observations (Gardner, 2000), this model proposed a range of inductive signals originating from the embryo and regulating trophectoderm development and implantation. Here, we investigate the role of signaling pathways forming the PrE under chemically-defined conditions, and its influence on post-implantation development of blastoids. Results Naïve pluripotency increase PrE differentiation.We used a high-content screening platform of non-adherent hydrogel microwells in 96 well-plates (Vrij et al., 2016a) to reproducibly aggregate small number of ESCs into EBs (Figure 1A). ESCs seeded into microwells were normally distributed across the 430 microwells within each well (7-12 cells per microwell) and formed EBs within 24 hours (Figure 1B, S1). We quantified PrE differentiation via in situ imaging of fluorescent reporters Pdgfrα (ESCs Pdgfrα-h2b-gfp/+ ) (Artus et al., 2010;Plusa et al., 2008) or Gata6 ESCs Gata6-h2b-venus/+ ). Leukemia inhibitory factor (Lif) appeared essential for the viability of EBs when formed with low cell numbers in serum-free B27N2 medium. However, EBs did not proliferate and formed only few PrE cells (1% and 4% of Pdgfrα+ EBs for 2D expansion in B27N2/2i/Lif and serum/Lif, respectively (Figure 1C). In contrast, serum/Lif induced proliferation and expression of Pdgfrα+ EBs (44%, Figure 1C, ...

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Section: Methodsmentioning

confidence: 99%

Section: Naïve Pluripotency Increase Pre Differentiationmentioning

confidence: 99%

Chemically-defined induction of a primitive endoderm and epiblast-like niche supports post-implantation progression from blastoids

et al. 2019

Preprint

View full text Add to dashboard Cite

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“…102 3D scaffolds that encapsulate ES cell-derived embryoid structures tune cell fate and germ layer specification by enabling the manipulation of cell-generated forces. 103 By mimicking the soft biophysical architecture of embryoid bodies early in development, artificial niches were used to guide iPSC self-organization to promote amniogenesis in a BMP-SMAD-dependent manner.…”

Section: Stem Cells Respond To Forcesmentioning

confidence: 99%

Mechanical forces direct stem cell behaviour in development and regeneration

Vining

Mooney

2017

Nat Rev Mol Cell Biol

1,177

958

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Stem cells and their local microenvironment, or niche, communicate through mechanical, cues to regulate cell fate and cell behaviour, and to guide developmental processes. During embryonic development, mechanical forces are involved in patterning and organogenesis. The physical environment of pluripotent stem cells regulates their differentiation and self-renewal. Mechanical and physical cues are also important in adult tissues, where adult stem cells require physical interactions with the extracellular matrix to maintain their potency. In vitro, synthetic models of the stem cell niche can be used to precisely control and manipulate the biophysical and biochemical properties of the stem cell microenvironment and examine how the mode and magnitude of mechanical cues, such as matrix stiffness or applied forces, direct stem cell differentiation and function. Fundamental insights on the mechanobiology of stem cells also inform the design of artificial niches to support stem cells for regenerative therapies.

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“…A GelMA hydrogel construct fabricated by casting in a PTFE mold was recently used for controlled release applications using an in vivo mouse model [37]. In addition, replica micromolding has become a standard technique to create low attachment microwell arrays to create cellular spheroids and micro-organoids, which can be used to drive bottom-up tissue engineering, to control stem cell behavior, and to improve drug screening [46–50]. …”

Section: Spatial Control Of Hydrogelsmentioning

confidence: 99%

Spatially and temporally controlled hydrogels for tissue engineering

Leijten

Seo

Yue

et al. 2017

Materials Science and Engineering: R: Reports

157

129

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Summary Recent years have seen tremendous advances in the field of hydrogel-based biomaterials. One of the most prominent revolutions in this field has been the integration of elements or techniques that enable spatial and temporal control over hydrogels’ properties and functions. Here, we critically review the emerging progress of spatiotemporal control over biomaterial properties towards the development of functional engineered tissue constructs. Specifically, we will highlight the main advances in the spatial control of biomaterials, such as surface modification, microfabrication, photo-patterning, and three-dimensional (3D) bioprinting, as well as advances in the temporal control of biomaterials, such as controlled release of molecules, photocleaving of proteins, and controlled hydrogel degradation. We believe that the development and integration of these techniques will drive the engineering of next-generation engineered tissues.

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Directed Assembly and Development of Material‐Free Tissues with Complex Architectures

Cited by 56 publications

References 45 publications

Chemically-defined induction of a primitive endoderm and epiblast-like niche supports post-implantation progression from blastoids

Chemically-defined induction of a primitive endoderm and epiblast-like niche supports post-implantation progression from blastoids

Mechanical forces direct stem cell behaviour in development and regeneration

Spatially and temporally controlled hydrogels for tissue engineering

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