Contributions of Mammalian Chimeras to Pluripotent Stem Cell Research.

Chimeras are widely acknowledged as the gold standard for assessing stem cell pluripotency, based on their capacity to test donor cell lineage potential in the context of an organized, normally developing tissue. Experimental chimeras provide key insights into mammalian developmental mechanisms and offer a resource for interrogating the fate potential of various pluripotent stem cell states. We highlight the applications and current limitations presented by intra- and inter-species chimeras and consider their future contribution to the stem cell field. Despite the technical and ethical demands of experimental chimeras, including human-interspecies chimeras, they are a provocative resource for achieving regenerative medicine goals.British Heart Foundation Centre of Regenerative Medicine, Wellcome Trust, Medical Research Council Cambridge Stem Cell Institute, Cambridge NIHR Biomedical Research CentreThis is the final version of the article. It first appeared from Elsevier via http://dx.doi.org/10.1016/j.stem.2016.07.01

Human-Mouse Chimerism Validates Human Stem Cell Pluripotency.

Pluripotent stem cells are defined by their capacity to differentiate into all three tissue layers that comprise the body. Chimera formation, generated by stem cell transplantation to the embryo, is a stringent assessment of stem cell pluripotency. However, the ability of human pluripotent stem cells (hPSCs) to form embryonic chimeras remains in question. Here we show using a stage-matching approach that human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) have the capacity to participate in normal mouse development when transplanted into gastrula-stage embryos, providing in vivo functional validation of hPSC pluripotency. hiPSCs and hESCs form interspecies chimeras with high efficiency, colonize the embryo in a manner predicted from classical developmental fate mapping, and differentiate into each of the three primary tissue layers. This faithful recapitulation of tissue-specific fate post-transplantation underscores the functional potential of hPSCs and provides evidence that human-mouse interspecies developmental competency can occur.This work was supported by National Institutes of Health grant No. 1R21ID012228 (R.A.P.); Medical Research Council/British Heart Foundation grant No. G1000847 (R.A.P.); British Heart Foundation Ph.D. studentship (V.L.M.); British Heart Foundation Centre of Regenerative Medicine (Oxford grant RM/13/3/3015); core support from the Wellcome Trust – Medical Research Council Cambridge Stem Cell Institute; and the Cambridge NIHR Biomedical Research Centre.This is the final version of the article. It first appeared from Cell Press via http://dx.doi.org/10.1016/j.stem.2015.11.01

Towards Large Scale Evaluation of Novel Sonification Techniques for Non Visual Shape Exploration

© 2015 ACM.There are several situations in which a person with visual impairment or blindness needs to extract information from an image. Examples include everyday activities, like reading a map, as well as educational activities, like exercises to develop visuospatial skills. In this contribution we propose a set of 6 sonification techniques to recognize simple shapes on touchscreen devices. The effectiveness of these sonification techniques is evaluated though Invisible Puzzle, a mobile application that makes it possible to conduct non-supervised evaluation sessions. Invisible Puzzle adopts a gamification approach and is a preliminary step in the development of a complete game that will make it possible to conduct a large scale evaluation with hundreds or thousands of blind users. With Invisible Puzzle we conducted 131 tests with sighted subjects and 18 tests with subjects with blindness. All subjects involved in the process successfully completed the evaluation session, with high engagement, hence showing the effectiveness of the evaluation procedure. Results give interesting insights on the differences among the sonification techniques and, most importantly, show that, after a short training, subjects are able to identify many different shapes

SafeBox : adaptable spatio-temporal generalization for location privacy protection

Spatial and temporal generalization emerged in the literature as a common approach to preserve location privacy. However, existing solutions have two main shortcomings. First, spatiotemporal generalization can be used with different objectives: for example, to guarantee anonymity or to decrease the sensitivity of the location information. Hence, the strategy used to compute the generalization can follow different semantics often depending on the privacy threat, while most of the existing solutions are specifically designed for a single semantics. Second, existing techniques prevent the so-called inversion attack by adopting a top-down strategy that needs to acquire a large amount of information. This may not be feasible when this information is dynamic (e.g., position or properties of objects) and needs to be acquired from external services (e.g., Google Maps). In this contribution we present a formal model of the problem that is compatible with most of the semantics proposed so far in the literature, and that supports new semantics as well. Our BottomUp algorithm for spatio-temporal generalization is compatible with the use of online services, it supports generalizations based on arbitrary semantics, and it is safe with respect to the inversion attack. By considering two datasets and two examples of semantics, we experimentally compare BottomUp with a more classical top-down algorithm, showing that BottomUp is efficient and guarantees better performance in terms of the average size (space and time) of the generalized regions

Gonio, aequus and incognitus : three spatial granularities for privacy-aware systems

Many solutions proposed in the literature to enforce privacy in presence of location information use, implicitly or explicitly, spatial granularities. However, most of the contributions do not describe the formal and computational properties of this tool in details. In this paper we propose three families of spatial granularities, specifically designed for privacy-aware systems. We mathematically characterize them and prove that all of them have two important formal properties. Then, for each of them, we show how to efficiently compute two fundamental operations

Robust derivation of epicardium and its differentiated smooth muscle cell progeny from human pluripotent stem cells.

The epicardium has emerged as a multipotent cardiovascular progenitor source with therapeutic potential for coronary smooth muscle cell, cardiac fibroblast (CF) and cardiomyocyte regeneration, owing to its fundamental role in heart development and its potential ability to initiate myocardial repair in injured adult tissues. Here, we describe a chemically defined method for generating epicardium and epicardium-derived smooth muscle cells (EPI-SMCs) and CFs from human pluripotent stem cells (HPSCs) through an intermediate lateral plate mesoderm (LM) stage. HPSCs were initially differentiated to LM in the presence of FGF2 and high levels of BMP4. The LM was robustly differentiated to an epicardial lineage by activation of WNT, BMP and retinoic acid signalling pathways. HPSC-derived epicardium displayed enhanced expression of epithelial- and epicardium-specific markers, exhibited morphological features comparable with human foetal epicardial explants and engrafted in the subepicardial space in vivo. The in vitro-derived epicardial cells underwent an epithelial-to-mesenchymal transition when treated with PDGF-BB and TGFβ1, resulting in vascular SMCs that displayed contractile ability in response to vasoconstrictors. Furthermore, the EPI-SMCs displayed low density lipoprotein uptake and effective lowering of lipoprotein levels upon treatment with statins, similar to primary human coronary artery SMCs. Cumulatively, these findings suggest that HPSC-derived epicardium and EPI-SMCs could serve as important tools for studying human cardiogenesis, and as a platform for vascular disease modelling and drug screening.This work was supported by the British Heart Foundation (BHF) [NH/11/1/28922], by the UK Medical Research Council and BHF [G1000847 to S.S. and R.A.P.), and by Cambridge Hospitals National Institute for Health Research Biomedical Research Centre funding (S.S. and R.A.P.). D.I. is on a University of Cambridge Commonwealth Scholarship. S.S. and F.S. are supported by the BHF [FS/13/29/ 30024]. L.G. is supported by the BHF [RM/l3/3/30159]. W.G.B. and V.L.M. are supported by BHF PhD studentships [FS/11/77/29327 and FS/10/48/28674].This is the final published version. It first appeared at http://dev.biologists.org/content/early/2015/03/25/dev.119271.abstract

Synchronized onset of nuclear and cell surface modifications in U937 cells during apoptosis

In this study we investigated the relationship between nuclear and cell surface modifications (i.e. blebbing, phosphatidylserine [PS] and sugar residues exposure) in a monocytic cell line, U937, during apoptosis induced by oxidative stress (1mM H2O2) or inhibition of protein synthesis (10 mg/ml puromycin). Dying cells were simultaneously observed for nuclear modifications, presence of superficial blebs and plasma membrane alterations. Morphological analysis performed by conventional fluorescence microscopy, or by transmission and scanning electron microscopy showed that the courses of nuclear and membrane alterations occured concomitantly, but the phenotype was dependent on the stage of the apoptotic process and the type of apoptogenic inducer used. The progression of apoptosis in U937 cells beyond early stages resulted in the extensive formation of blebs which concomitantly lost some typical markers of apoptosis, such as PS and sugar residues. Therefore, the modality by which the nucleus condenses, or the amount and the pattern of distribution of PS on the cell surface were, for each cell line, strictly related to the apoptogenic inducer. The morphological data reported in the present paper should lead to a more precise quantification of apoptosis by improving the detection of apoptotic cells in vivo (i.e. in tissue, organs), which is a crucial point in the evaluation of efficiency of antiproliferative drugs, such as antiblastic or immunosuppressive compounds

The NATO project : nanoparticle-based countermeasures for microgravity-induced osteoporosis

Recent advances in nanotechnology applied to medicine and regenerative medicine have an enormous and unexploited potential for future space and terrestrial medical applications. The Nanoparticles and Osteoporosis (NATO) project aimed to develop innovative countermeasures for secondary osteoporosis affecting astronauts after prolonged periods in space microgravity. Calcium- and Strontium-containing hydroxyapatite nanoparticles (nCa-HAP and nSr-HAP, respectively) were previously developed and chemically characterized. This study constitutes the first investigation of the effect of the exogenous addition of nCa-HAP and nSr-HAP on bone remodeling in gravity (1\u2009g), Random Positioning Machine (RPM) and onboard International Space Station (ISS) using human bone marrow mesenchymal stem cells (hBMMSCs). In 1\u2009g conditions, nSr-HAP accelerated and improved the commitment of cells to differentiate towards osteoblasts, as shown by the augmented alkaline phosphatase (ALP) activity and the up-regulation of the expression of bone marker genes, supporting the increased extracellular bone matrix deposition and mineralization. The nSr-HAP treatment exerted a protective effect on the microgravity-induced reduction of ALP activity in RPM samples, and a promoting effect on the deposition of hydroxyapatite crystals in either ISS or 1\u2009g samples. The results indicate the exogenous addition of nSr-HAP could be potentially used to deliver Sr to bone tissue and promote its regeneration, as component of bone substitute synthetic materials and additive for pharmaceutical preparation or food supplementary for systemic distribution