In vitro organogenesis in three dimensions: self-organising stem cells

Sasai, Yoshiki; Eiraku, Mototsugu; Suga, Hidetaka

doi:10.1242/dev.079590

Cited by 182 publications

(124 citation statements)

References 106 publications

(111 reference statements)

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“…Remarkable epithelial morphogenesis was recapitulated in the neo‐tissues formed in the cell‐laden THAG, where Caco2 and Hep G2 cells developed into tubal organoids ( Figure 5 a,c, Figures S8 and S9, Supporting Information) 35, 36. IF co‐staining for human retinol‐binding protein II (RBP 2) or human hepatic nuclear factor 4 alpha (HNF 4α)—markers of Caco2 and Hep G2 cells,34, 37 respectively—and E‐cadherin further testified that the tubular structures were constituted by the formed epithelium organoids (Figure 5b and d).…”

Section: Resultsmentioning

confidence: 99%

Engineering a Tumor Microenvironment‐Mimetic Niche for Tissue Regeneration with Xenogeneic Cancer Cells

Wang

Abudukeremu

et al. 2018

Advanced Science

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The insufficient number of cells suitable for transplantation is a long‐standing problem to cell‐based therapies aimed at tissue regeneration. Xenogeneic cancer cells (XCC) may be an alternative source of therapeutic cells, but their transplantation risks both immune rejection and unwanted spreading. In this study, a strategy to facilitate XCC transplantation is reported and their spreading in vivo is confined by constructing an engineering matrix that mimics the characteristics of tumor microenvironment. The data show that this matrix, a tumor homogenate‐containing hydrogel (THAG), successfully creates an immunosuppressive enclave after transplantation into immunocompetent mice. XCC of different species and tissue origins seeded into THAG survive well, integrated with the host and developed the intrinsic morphology of the native tissue, without being eliminated or spreading out of the enclave. Most strikingly, immortalized human hepatocyte cells and rat β‐cells loaded into THAG exert the physiological functions of the human liver and rat pancreas islets, respectively, in the mouse body. This study demonstrates a novel and feasible approach to harness the unique features of tumor development for tissue transplantation and regenerative medicine.

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Section: Resultsmentioning

confidence: 99%

Engineering a Tumor Microenvironment‐Mimetic Niche for Tissue Regeneration with Xenogeneic Cancer Cells

Wang

Abudukeremu

et al. 2018

Advanced Science

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“…However, reproducibly recapitulating symmetry breaking in vitro has proven to be challenging. Indeed, in making the optic cup in vitro, despite exhibiting striking similarities to in vivo cell organization, the organoid still lacked some asymmetric aspects, such as the gapped structure at the ventralmost region of the optic cup seen in the embryo (Sasai et al, 2012). In addition, the spontaneous reaggregation of chick embryo cells in sorting experiments, although resulting in the correct placement of cell types relative to one another, resulted in a radially symmetric pattern, despite these cells not having a radially symmetric pattern in the embryo (Steinberg, 1963).…”

Section: Breaking Symmetrymentioning

confidence: 99%

“…This structure, termed an organoid (see Glossary, Box 1), exhibits strong similarities to the in vivo tissue; it underwent changes in local tissue mechanics, invaginating to form the characteristic morphology of the optical cup ( Fig. 1A) (Eiraku et al, 2011;Sasai et al, 2012). Importantly, this organoid formed without external scaffolding or mechanics, further demonstrating the innate ability of cells to self-generate functional multicellular structures.…”

Section: Introductionmentioning

confidence: 97%

“…In general, an organoid is defined as a structure in which pluripotent or progenitor stem cells are differentiated into multiple cell populations that self-organize/assemble into a tissue that resembles an organ in vivo (Clevers, 2016;Fatehullah et al, 2016;Kicheva and Briscoe, 2015;Lancaster and Knoblich, 2014;Sasai et al, 2012;Turner et al, 2016). In a similar vein, researchers have also generated structures referred to as embryoid bodies, embryoids and gastruloids (see Glossary, Box 1; Fig.…”

Section: Introductionmentioning

confidence: 99%

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Embryoids, organoids and gastruloids: new approaches to understanding embryogenesis

2017

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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.

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“…Organoid protocols stand at the forefront of these technologies, as these 3D approaches more accurately reproduce in vivo developmental events leading to more precise in vitro models 2,3 . Organoids have already been developed for several organ systems, including retina 4 , intestine 5 , thyroid 6 , liver 7 , pituitary 8 , inner ear 9 , kidney 10-12 and brain 13 .…”

Section: Introductionmentioning

confidence: 99%

Generation of cerebral organoids from human pluripotent stem cells

2014

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protocol INtroDUctIoNIn vitro methods to model human development and disease are part of a rapidly expanding field of stem cell biology with major therapeutic implications 1 . Organoid protocols stand at the forefront of these technologies, as these 3D approaches more accurately reproduce in vivo developmental events leading to more precise in vitro models 2,3 . Organoids have already been developed for several organ systems, including retina 4 , intestine 5 , thyroid 6 , liver 7 , pituitary 8 , inner ear 9 , kidney 10-12 and brain 13 . However, only a handful of methods exist for generating such tissues from human pluripotent stem cells (hPSCs) [13][14][15][16][17] . In this protocol, we describe the methodology that was recently published for generating brain organoid tissues from hPSCs 13 . Development of the protocolThe method described here builds on an extensive foundation of protocols for neural differentiation, 3D tissue culture and tissue engineering. Cerebral organoids develop through intrinsic selforganizing processes upon timely application of components and culture environments that had previously been described individually. Thus, the method is an amalgamation of previous methods, combined in a specific manner to address two main objectives: (i) the establishment of neural identity and differentiation and (ii) the recapitulation of 3D structural organization. Establishment of brain identityThe first goal of the protocol is induction and differentiation of neural tissue. This involves identification of media formulations and additives to drive neural identity and to stimulate brain development in vitro. To achieve this, a number of medium formulations were tested at various time points. Rather than describing the multitude of tested combinations, we focus here on the successful outcome.Neural tissue develops in vivo from a germ layer called the ectoderm 18 . Similarly, PSCs in vitro can be stimulated to develop germ layers, including ectoderm, within aggregates called embryoid bodies (EBs) 19 . A number of previous studies have described successful differentiation of EBs in embryonic stem cell (ESC) medium with decreased basic fibroblast growth factor (bFGF) 20 and high-dose rho-associated protein kinase (ROCK) inhibitor to limit cell death 21 . Similarly, cerebral organoids develop from EBs grown initially in ESC medium with low bFGF and ROCK inhibitor.Subsequent neural induction of EBs follows a minimal medium formulation very similar to that established by Zhang et al. 22,23 for the induction of neural rosettes, a 2D polarized organization of neuroepithelial cells. However, for the generation of cerebral organoids, EBs are kept in suspension, leading to uniform neural ectoderm formation along the outer surface of EBs, whereas inner non-neural mesendodermal tissues do not develop.Neural ectoderm in vivo establishes radially organized neuroepithelia that expand to form various brain structures. Similarly, organoids placed in a differentiation medium that supports both neural progenitors and their p...

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In vitro organogenesis in three dimensions: self-organising stem cells

Cited by 182 publications

References 106 publications

Engineering a Tumor Microenvironment‐Mimetic Niche for Tissue Regeneration with Xenogeneic Cancer Cells

Engineering a Tumor Microenvironment‐Mimetic Niche for Tissue Regeneration with Xenogeneic Cancer Cells

Embryoids, organoids and gastruloids: new approaches to understanding embryogenesis

Generation of cerebral organoids from human pluripotent stem cells

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