Data describing Rax positive optic-vesicle generation from mouse embryonic stem cells in vitro

AbstractThis article contains data related to the research article entitled “Specification of embryonic stem cell-derived tissues into eye fields by Wnt signaling using rostral diencephalic tissue-inducing culture” Sakakura (2016) [1]. Mouse embryonic stem cells (ESC) were used for the generation of optic vesicle-like tissues in vitro. In this article we described data in which a Rax::GFP knock-in ESC line was used to monitor the formation of optic tissues. In addition, we also described the data of regional marker expression of Rax, Sox2 and Pax6 in vivo around the forebrain and the eye tissues for comparative purposes. These data can be valuable to researchers interested in investigating forebrain and eye tissue development

How might we build limbs in vitro informed by the modular aspects and tissue-dependency in limb development?

Building limb morphogenesis in vitro would substantially open up avenues for research and applications of appendage development. Recently, advances in stem cell engineering to differentiate desired cell types and produce multicellular structures in vitro have enabled the derivation of limb-like tissues from pluripotent stem cells. However, in vitro recapitulation of limb morphogenesis is yet to be achieved. To formulate a method of building limbs in vitro, it is critically important to understand developmental mechanisms, especially the modularity and the dependency of limb development on the external tissues, as those would help us to postulate what can be self-organized and what needs to be externally manipulated when reconstructing limb development in vitro. Although limbs are formed on the designated limb field on the flank of embryo in the normal developmental context, limbs can also be regenerated on the amputated stump in some animals and experimentally induced at ectopic locations, which highlights the modular aspects of limb morphogenesis. The forelimb-hindlimb identity and the dorsal-ventral, proximal-distal, and anterior-posterior axes are initially instructed by the body axis of the embryo, and maintained in the limb domain once established. In contrast, the aspects of dependency on the external tissues are especially underscored by the contribution of incoming tissues, such as muscles, blood vessels, and peripheral nerves, to developing limbs. Together, those developmental mechanisms explain how limb-like tissues could be derived from pluripotent stem cells. Prospectively, the higher complexity of limb morphologies is expected to be recapitulated by introducing the morphogen gradient and the incoming tissues in the culture environment. Those technological developments would dramatically enhance experimental accessibility and manipulability for elucidating the mechanisms of limb morphogenesis and interspecies differences. Furthermore, if human limb development can be modeled, drug development would be benefited by in vitro assessment of prenatal toxicity on congenital limb deficiencies. Ultimately, we might even create a future in which the lost appendage would be recovered by transplanting artificially grown human limbs

Dendritic cells cross talk with tumor antigen-specific CD8+T cells, Vγ9γδT cells, and Vα24NKT cells in patients with glioblastoma multiforme and in healthy donors

The finding that dendritic cells (DCs) orchestrate innate and adaptive immune responses has stimulated research on harnessing DCs for developing more effective vaccines for DC therapy. The expression of cytomegalovirus (CMV) antigens in glioblastoma multiforme (GBM) presents a unique opportunity to target these viral proteins for tumor immunotherapy. Here, we demonstrate that Vγ9γδT cells, innate immune cells activated by zoledronate (Z), and Vα24NKT cells, innate/adaptive immune cells activated by α-galactosylceramide (G) can link innate and adaptive immunities through cross talk with IFN-DCs from patients with GBM and healthy donors in a way that can amplify the activation and proliferation of CMVpp65-specific CD8+T cells. The IFN-DCs derived from patients with GBM used in this study express lower levels of programmed death ligand (PDL)1 and PDL2 and higher levels of CCR7 than the most commonly used mature IL-4DCs. The expression level of programmed cell death 1 (PD1) on CD8+ T cells, including CMVpp65-specific CD8+T cells, expanded by IFN-DCs pulsed with the CMVpp65-peptide and Z plus G (IFN-DCs/P+Z+G) was lower than that expanded by IFN-DCs pulsed with the peptide alone (IFN-DCs/P). Multifunctional T cells, including HLA-A*0201-restricted CMVpp65-specific CD8+T cells, Vγ9γδT cells, and Vα24NKT cells, efficiently kill HLA-A*0201 positive GBM cell line expressing CMVpp65 protein (T98G). These findings indicate that DC therapy using IFN-DCs/P+Z+G and/or CTL therapy using CMVpp65-specific CD8+T cells expanded by IFN-DCs/P+Z+G may lead to a good clinical outcome for patients with GBM. This article is protected by copyright. All rights reserved

Self-Organized Formation of Polarized Cortical Tissues from ESCs and Its Active Manipulation by Extrinsic Signals

SummaryHere, we demonstrate self-organized formation of apico-basally polarized cortical tissues from ESCs using an efficient three-dimensional aggregation culture (SFEBq culture). The generated cortical neurons are functional, transplantable, and capable of forming proper long-range connections in vivo and in vitro. The regional identity of the generated pallial tissues can be selectively controlled (into olfactory bulb, rostral and caudal cortices, hem, and choroid plexus) by secreted patterning factors such as Fgf, Wnt, and BMP. In addition, the in vivo-mimicking birth order of distinct cortical neurons permits the selective generation of particular layer-specific neurons by timed induction of cell-cycle exit. Importantly, cortical tissues generated from mouse and human ESCs form a self-organized structure that includes four distinct zones (ventricular, early and late cortical-plate, and Cajal-Retzius cell zones) along the apico-basal direction. Thus, spatial and temporal aspects of early corticogenesis are recapitulated and can be manipulated in this ESC culture

Remodeling of Monoplanar Purkinje Cell Dendrites during Cerebellar Circuit Formation

Dendrite arborization patterns are critical determinants of neuronal connectivity and integration. Planar and highly branched dendrites of the cerebellar Purkinje cell receive specific topographical projections from two major afferent pathways; a single climbing fiber axon from the inferior olive that extend along Purkinje dendrites, and parallel fiber axons of granule cells that contact vertically to the plane of dendrites. It has been believed that murine Purkinje cell dendrites extend in a single parasagittal plane in the molecular layer after the cell polarity is determined during the early postnatal development. By three-dimensional confocal analysis of growing Purkinje cells, we observed that mouse Purkinje cells underwent dynamic dendritic remodeling during circuit maturation in the third postnatal week. After dendrites were polarized and flattened in the early second postnatal week, dendritic arbors gradually expanded in multiple sagittal planes in the molecular layer by intensive growth and branching by the third postnatal week. Dendrites then became confined to a single plane in the fourth postnatal week. Multiplanar Purkinje cells in the third week were often associated by ectopic climbing fibers innervating nearby Purkinje cells in distinct sagittal planes. The mature monoplanar arborization was disrupted in mutant mice with abnormal Purkinje cell connectivity and motor discoordination. The dendrite remodeling was also impaired by pharmacological disruption of normal afferent activity during the second or third postnatal week. Our results suggest that the monoplanar arborization of Purkinje cells is coupled with functional development of the cerebellar circuitry

The bending of cell sheets - from folding to rolling

The bending of cell sheets plays a major role in multicellular embryonic morphogenesis. Recent advances are leading to a deeper understanding of how the biophysical properties and the force-producing behaviors of cells are regulated, and how these forces are integrated across cell sheets during bending. We review work that shows that the dynamic balance of apical versus basolateral cortical tension controls specific aspects of invagination of epithelial sheets, and recent evidence that tissue expansion by growth contributes to neural retinal invagination in a stem cell-derived, self-organizing system. Of special interest is the detailed analysis of the type B inversion in Volvox reported in BMC Biology by Höhn and Hallmann, as this is a system that promises to be particularly instructive in understanding morphogenesis of any monolayered spheroid system

Coordinated Morphogenetic Mechanisms Shape the Vertebrate Eye.

The molecular bases of vertebrate eye formation have been extensively investigated during the past 20 years. This has resulted in the definition of the backbone of the gene regulatory networks controlling the different steps of eye development and has further highlighted a substantial conservation of these networks among vertebrates. Yet, the precise morphogenetic events allowing the formation of the optic cup from a small group of cells within the anterior neural plate are still poorly understood. It is also unclear if the morphogenetic events leading to eyes of very similar shape are indeed comparable among all vertebrates or if there are any species-specific peculiarities. Improved imaging techniques have enabled to follow how the eye forms in living embryos of a few vertebrate models, whereas the development of organoid cultures has provided fascinating tools to recapitulate tissue morphogenesis of other less accessible species. Here, we will discuss what these advances have taught us about eye morphogenesis, underscoring possible similarities and differences among vertebrates. We will also discuss the contribution of cell shape changes to this process and how morphogenetic and patterning mechanisms integrate to assemble the final architecture of the eye

Symptomatic Developmental Venous Anomaly with an Increased β2-microglobulin Level in Cerebrospinal Fluid: A Case Report

Background: Gadolinium-enhanced magnetic resonance imaging (MRI) can be used to observe the progression of cerebral infarction, which sometimes mimics malignant brain tumors. While the β2-microglobulin (β2MG) level in blood plasma or cerebrospinal fluid (CSF) is useful for the diagnosis of malignant tumors or degenerative diseases, these results may create confusion regarding a definitive diagnosis, because it is not a specific marker. We present a rare case of symptomatic developmental venous anomaly (DVA), accompanied by transient, irregular, enhanced cerebral lesions and elevated β2MG in the CSF. Case Description: A 56-year-old woman developed dysarthria and underwent MRI, which revealed a right frontal hyperintense area around a previous lesion on diffusion-weighted imaging (DWI). She was treated based on the tentative diagnosis of an ischemic cerebrovascular event, and symptoms subsided in 3 days. MRI on day 7 revealed an enlargement of the hyperintense area on DWI. Post-gadolinium MRI showed multiple, enhanced patchy areas in the right frontal lobe and an abnormally large vein connected to dilated medullary venules, indicating DVA. Magnetic resonance angiography showed no stenosis or arterial occlusion. The β2MG level in the CSF was elevated at 2,061 μg/l, and a differential diagnosis from malignant tumor was required. However, MRI on day 23 revealed total disappearance of the enhanced lesions and a decrease in the high intensity area on DWI. Considering the clinical course, the DVA was symptomatic because of the perfusion disturbance. Conclusion: Careful evaluation is necessary when considering the associated pathologies and potential complications of DVA if detected near a gadolinium-enhanced lesion

Opportunities for organoids as new models of aging.

The biology of aging is challenging to study, particularly in humans. As a result, model organisms are used to approximate the physiological context of aging in humans. However, the best model organisms remain expensive and time-consuming to use. More importantly, they may not reflect directly on the process of aging in people. Human cell culture provides an alternative, but many functional signs of aging occur at the level of tissues rather than cells and are therefore not readily apparent in traditional cell culture models. Organoids have the potential to effectively balance between the strengths and weaknesses of traditional models of aging. They have sufficient complexity to capture relevant signs of aging at the molecular, cellular, and tissue levels, while presenting an experimentally tractable alternative to animal studies. Organoid systems have been developed to model many human tissues and diseases. Here we provide a perspective on the potential for organoids to serve as models for aging and describe how current organoid techniques could be applied to aging research