Formation of Spatially and Geometrically Controlled Three-Dimensional Tissues in Soft Gels by Sacrificial Micromolding

Cerchiari, Alec E.; Garbe, James C.; Todhunter, Michael E.; Jee, Noel Y.; Pinney, James R.; LaBarge, Mark A.; Desai, Tejal A.; Gartner, Zev J.

doi:10.1089/ten.tec.2014.0450

Cited by 23 publications

(30 citation statements)

References 31 publications

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“…These purified populations of cultured primary cells were reconstituted by chemical or mechanical means at 50:50 ratios, either fully embedded within Matrigel or in nonfouling microwells (agarose) (Fig. 1B and SI Appendix) (20). Consistent with their ability to self-organize, LEP and MEP efficiently formed the correct architecture in Matrigel after 12-24 h (Movie S1).…”

Section: Human Mammary Epithelial Cells Can Self-organize Into An Invmentioning

confidence: 95%

A strategy for tissue self-organization that is robust to cellular heterogeneity and plasticity

Cerchiari

Garbe

Jee³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Developing tissues contain motile populations of cells that can selforganize into spatially ordered tissues based on differences in their interfacial surface energies. However, it is unclear how self-organization by this mechanism remains robust when interfacial energies become heterogeneous in either time or space. The ducts and acini of the human mammary gland are prototypical heterogeneous and dynamic tissues comprising two concentrically arranged cell types. To investigate the consequences of cellular heterogeneity and plasticity on cell positioning in the mammary gland, we reconstituted its selforganization from aggregates of primary cells in vitro. We find that self-organization is dominated by the interfacial energy of the tissue-ECM boundary, rather than by differential homo-and heterotypic energies of cell-cell interaction. Surprisingly, interactions with the tissue-ECM boundary are binary, in that only one cell type interacts appreciably with the boundary. Using mathematical modeling and cell-type-specific knockdown of key regulators of cell-cell cohesion, we show that this strategy of self-organization is robust to severe perturbations affecting cell-cell contact formation. We also find that this mechanism of self-organization is conserved in the human prostate. Therefore, a binary interfacial interaction with the tissue boundary provides a flexible and generalizable strategy for forming and maintaining the structure of two-component tissues that exhibit abundant heterogeneity and plasticity. Our model also predicts that mutations affecting binary cell-ECM interactions are catastrophic and could contribute to loss of tissue architecture in diseases such as breast cancer.heterogeneity | cell sorting | differential adhesion | mammary | prostate S elf-organization is a process that contributes to pattern formation and repair at all scales of biological complexity. At the tissue scale, defining robust strategies of self-organization is critical for engineering functional tissues, as well as for understanding development and the breakdown of tissue structure during diseases such as cancer (1). During development, two or more populations of motile cells can self-organize into spatially ordered tissues by a process referred to as cell sorting (2-4). The outcome of cell sorting can be rationalized using physical models that invoke cell-type-specific differences in interfacial energies. Interfacial energies arise through the action of a contractile cell cortex coupled to adhesion molecules (e.g., cadherins) that link the cortices of neighboring cells and signal to modulate cortical tension at specific cellular interfaces (5). In general, the organization of a tissue after cell sorting corresponds to a configuration that maximizes the formation of the most energetically favorable (hereafter referred to as most cell-cell cohesive) † cellular interfaces (6). For example, with an intermediate level of heterotypic cell-cell cohesion the most self-cohesive cell type is typically found in the tissue core, with the le...

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Section: Human Mammary Epithelial Cells Can Self-organize Into An Invmentioning

confidence: 95%

A strategy for tissue self-organization that is robust to cellular heterogeneity and plasticity

Cerchiari

Garbe

Jee³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

111

120

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“…Based on our analytical model for tissue self-organization 10 , we reasoned that the higher affinity of epithelial cells for adhesive ECM relative to a non-adhesive microparticle could direct an aggregate of cells to push a microparticle into their luminal space when fully embedded in adhesive ECM. We previously demonstrated a technique called Sacrifical Micromolding that allows collections of 10 to 50 epithelial cells to be aggregated, fully embedded in biomimetic hydrogels such as Matrigel or collagen, and self-organize into a lumenized cyst 11 . We therefore used Sacrificial Micromolding 10 to pre-aggregate Caco-2 cells and polymeric microparticles within degradable microwells, before transferring these aggregates to Matrigel culture.…”

Section: Resultsmentioning

confidence: 99%

Probing the luminal microenvironment of reconstituted epithelial microtissues

Cerchiari

Samy

Todhunter

et al. 2016

Sci Rep

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Polymeric microparticles can serve as carriers or sensors to instruct or characterize tissue biology. However, incorporating microparticles into tissues for in vitro assays remains a challenge. We exploit three-dimensional cell-patterning technologies and directed epithelial self-organization to deliver microparticles to the lumen of reconstituted human intestinal microtissues. We also develop a novel pH-sensitive microsensor that can measure the luminal pH of reconstituted epithelial microtissues. These studies offer a novel approach for investigating luminal microenvironments and drug-delivery across epithelial barriers.

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“…This technique also facilitates the production of organoids with more complicated shapes, such as branching patterns. For softer ECMs that cannot be directly stamped, Cerchiari et al (2015b) developed a technique using gelatin as a degradable scaffold to produce microwells that could then be removed by buffer exchange and replaced by matrices such as Matrigel or fibrin, maintaining the positional fidelity of the original wells. Both of these latter methods allow the production of organoids of non-spherical shapes that can be used to dissect how morphogen gradients are set up and maintained in the stem cell niche.…”

Section: Microwells Provide Control Over Organoid Shape and Sizementioning

confidence: 99%

Dissecting the stem cell niche with organoid models: an engineering-based approach

2017

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For many tissues, single resident stem cells grown in vitro under appropriate three-dimensional conditions can produce outgrowths known as organoids. These tissues recapitulate much of the cell composition and architecture of the in vivo organ from which they derive, including the formation of a stem cell niche. This has facilitated the systematic experimental manipulation and single-cell, highthroughput imaging of stem cells within their respective niches. Furthermore, emerging technologies now make it possible to engineer organoids from purified cellular and extracellular components to directly model and test stem cell-niche interactions. In this Review, we discuss how organoids have been used to identify and characterize stem cell-niche interactions and uncover new niche components, focusing on three adult-derived organoid systems. We also describe new approaches to reconstitute organoids from purified cellular components, and discuss how this technology can help to address fundamental questions about the adult stem cell niche.

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Formation of Spatially and Geometrically Controlled Three-Dimensional Tissues in Soft Gels by Sacrificial Micromolding

Cited by 23 publications

References 31 publications

A strategy for tissue self-organization that is robust to cellular heterogeneity and plasticity

A strategy for tissue self-organization that is robust to cellular heterogeneity and plasticity

Probing the luminal microenvironment of reconstituted epithelial microtissues

Dissecting the stem cell niche with organoid models: an engineering-based approach

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