Lineage Commitment of Conditionally Immortalized Bone Marrow Mesenchymal Stromal Cells from Tetracycline-Regulated SV40 Large T-antigen Transgenic Mice

Adult bone marrow contains a population of mesenchymal stem cells capable to self-renew and to differentiate into haematopoietic-supportive stroma, osteo, adipo- and chondrocytes. However, the identity of mesenchymal stem cells still remains uncertain. The complex population of their descendants, bone marrow mesenchymal stromal cells (BM MSCs), represents a model to study the principles of differentiation and commitment into mesodermal lineages. The experiments using BM MSCs are often hampered by their low proliferative capacity in vitro. In the present study, we established conditionally immortalized BM MSCs from tetracycline-regulated SV40 Large T-antigen transgenic mice. The identity of the conditionally immortalized BM MSCs was confirmed by marker expression, ability to support haematopoiesis and differentiation potential. The advantages of the conditional immortalization are encompassed in (1) indefinite expansion of cell populations, (2) possibility to perform cellular cloning and (3) prevention from spontaneous differentiation. We demonstrated the heterogeneity of BM MSCs and identified at least 6 types of progenitors within BM MSCs population based on their differentiation potential (“OAC”, “OA”, “OC”, “AC”, “O”, “A”). A hypothetical model of BM MSC hierarchy and the relationships between the progenitors has been proposed. We observed that the Wnt/β-catenin signaling pathway and GSK3 activity could modulate the efficiency of osteo- and adipogenic differentiation pathways, but we didn’t find evidence that the lineage commitment of BM MSCs is determined by Wnt. We elucidated the mechanism of transcriptional regulation of the adipogenic induction of BM MSCs in vitro. Our data revealed the key regulatory role of PPARγ1 during adipogenesis in BM MSCs. Furthermore, we assume that PPARγ1 is a potential trigger of the adipogenic commitment of the BM MSCs progenitors. Finally, the non-adipogenic BM MSCs progenitors were converted into the adipogenic lineage using ectopical expression of the transcription factors C/EBPα, C/EBPβ and C/EBPδ. Our findings provide a novel insight into the molecular mechanisms of BM MSCs lineage commitment

Towards consistent generation of pancreatic lineage progenitors from human pluripotent stem cells.

Human pluripotent stem cells can in principle be used as a source of any differentiated cell type for disease modelling, drug screening, toxicology testing or cell replacement therapy. Type I diabetes is considered a major target for stem cell applications due to the shortage of primary human beta cells. Several protocols have been reported for generating pancreatic progenitors by in vitro differentiation of human pluripotent stem cells. Here we first assessed one of these protocols on a panel of pluripotent stem cell lines for capacity to engender glucose sensitive insulin-producing cells after engraftment in immunocompromised mice. We observed variable outcomes with only one cell line showing a low level of glucose response. We, therefore, undertook a systematic comparison of different methods for inducing definitive endoderm and subsequently pancreatic differentiation. Of several protocols tested, we identified a combined approach that robustly generated pancreatic progenitors in vitro from both embryo-derived and induced pluripotent stem cells. These findings suggest that, although there are intrinsic differences in lineage specification propensity between pluripotent stem cell lines, optimal differentiation procedures may consistently direct a substantial fraction of cells into pancreatic specification.This research was supported by European Commission Grant agreement 241883, “BetaCellTherapy”, and by the United Kingdom Medical Research Council.This is the final version of the article. It first appeared from Royal Society Publishing via http://dx.doi.org/10.1098/rstb.2014.036

Capacitation of human naïve pluripotent stem cells for multi-lineage differentiation.

Human naïve pluripotent stem cells (PSCs) share features with the pre-implantation epiblast. They therefore provide an unmatched opportunity for characterising the developmental programme of pluripotency in Homo sapiens Here, we confirm that naïve PSCs do not respond directly to germ layer induction, but must first acquire competence. Capacitation for multi-lineage differentiation occurs without exogenous growth factor stimulation and is facilitated by inhibition of Wnt signalling. Whole-transcriptome profiling during this formative transition highlights dynamic changes in gene expression, which affect many cellular properties including metabolism and epithelial features. Notably, naïve pluripotency factors are exchanged for postimplantation factors, but competent cells remain devoid of lineage-specific transcription. The gradual pace of transition for human naïve PSCs is consistent with the timespan of primate development from blastocyst to gastrulation. Transcriptome trajectory during in vitro capacitation of human naïve cells tracks the progression of the epiblast during embryogenesis in Macaca fascicularis, but shows greater divergence from mouse development. Thus, the formative transition of naïve PSCs in a simple culture system may recapitulate essential and specific features of pluripotency dynamics during an inaccessible period of human embryogenesis.This research was funded by the Medical Research Council of the United Kingdom (G1001028 and MR/P00072X/1), the European Commission Framework 7 (HEALTH-F4-2013-602423, PluriMes) and the UK Regenerative Medicine Platform (MR/L012537/1). The Cambridge Stem Cell Institute receives core funding from the Wellcome Trust and the Medical Research Council. AS is a Medical Research Council Professor

On-Stage Playing Space As a Resource of Managers’ Professional Development

The article is devoted to the activities of the Laboratory “Workshop stage action in the management” of the Department of control theory on the formation stage methodology in management education. Appeal to this modern practice gives the Manager a chance to be holistic in action and to work effectively in a modern crisis situation. The stage space of the game becomes the site of formation of organizational and managerial positions in a situation of uncertainty and learning resource is an organic action of one’s own physicality (body and voice). Stage methodology opens up a different horizon of management education towards organic management

The challenge of social innovation : approaches and key mechanisms of development

Recently, a key motive for innovation has been the generation of economic value. Currently we are facing a challenge to develop new approaches to involving the public in solving social problems through innovation based on collaboration and cooperation. Consequently, there is an urgent requirement to shape a favorable environment for innovation, creating both economic and social value. The purpose of this study is to reveal mechanisms for the development of social innovation that can be successfully introduced and implemented in Russia. The advantage of a systems-based approach to social innovation is that social innovation is defined as institutional change leading to the emergence of new routines (traditions) or practices. The use of benchmarking, along with comparative and historical analysis, to study foreign experiences of social innovation makes it possible to identify best practice in creating the conditions needed to develop social innovations, organize innovation processes and promote systemic innovations. On the basis of information received, key mechanisms of social innovation were identified, including that of innovation mediation. The system capabilities of Living Labs in the promotion of social innovations were, in particular, investigated.peer-reviewe

Moscow youth non-governmental organizations as a youth behavior regulator

The Russian youth of today are inhomogeneous in terms of their composition, deeply polarized as to their social, national and ethnic characteristics and have unequal opportunities for starting their lives and careers.   The objective of this research is to identify the preventive influence of youth non-governmental organizations on the illegal activities of students and reduction of the manifestations of destructive behavior patterns. The obtained results have shown that the general mood of participants of youth non-governmental organizations consists in preservation of the identity of each non-governmental organization, defining it as a unique establishment with its own elaborated solutions and effectiveness. It has been found that the main focus areas of Moscow youth non-governmental organizations are very diverse and embrace nearly all spheres of social life

The ploidy and genetic structure of hybrid population of water frogs Pelophylax esculentus complex (Amphibia, Ranidae) of Ukraine fauna

The complex study, including allozyme variability and cytometry of hybrid populations of green frogs Pelophylax esculentus (L., 1758) complex has confirmed that the only region of Ukraine where allodip loid are encountered frequently is the Severski Donets basin (9% of all hybrids). In other areas, only two poly ploidy hybrids (0.9%) and one probably autopolyploid individual of each parental species have been regis tered. According to allozyme specters, all three polyploidy hybrids from the Severski Donets basin were males and belonged to biotype P. esculentus (=lessonae) – 2 ridibundus, and their population in this region has halved during the past decade

Differential Expression of Surface Markers in Mouse Bone Marrow Mesenchymal Stromal Cell Subpopulations with Distinct Lineage Commitment

Bone marrow mesenchymal stromal cells (BM MSCs) represent a heterogeneous population of progenitors with potential for generation of skeletal tissues. However the identity of BM MSC subpopulations is poorly defined mainly due to the absence of specific markers allowing in situ localization of those cells and isolation of pure cell types. Here, we aimed at characterization of surface markers in mouse BM MSCs and in their subsets with distinct differentiation potential. Using conditionally immortalized BM MSCs we performed a screening with 176 antibodies and high-throughput flow cytometry, and found 33 markers expressed in MSCs, and among them 3 were novel for MSCs and 13 have not been reported for MSCs from mice. Furthermore, we obtained clonally derived MSC subpopulations and identified bipotential progenitors capable for osteo- and adipogenic differentiation, as well as monopotential osteogenic and adipogenic clones, and thus confirmed heterogeneity of MSCs. We found that expression of CD200 was characteristic for the clones with osteogenic potential, whereas SSEA4 marked adipogenic progenitors lacking osteogenic capacity, and CD140a was expressed in adipogenic cells independently of their efficiency for osteogenesis. We confirmed our observations in cell sorting experiments and further investigated the expression of those markers during the course of differentiation. Thus, our findings provide to our knowledge the most comprehensive characterization of surface antigens expression in mouse BM MSCs to date, and suggest CD200, SSEA4 and CD140a as markers differentially expressed in distinct types of MSC progenitors

Pathophysiological mechanisms of renal damage in obstructive uropathies as potential therapeutic targets: A literature review

Obstructive uropathies are a group of conditions characterized by urinary tract blockages, leading to impaired urine flow and renal damage. This comprehensive literature review aims to explore the pathophysiological mechanisms underlying renal damage in obstructive uropathies and identify potential therapeutic targets for intervention. The review synthesizes current knowledge from a wide range of studies and provides an overview of the complex cellular and molecular processes involved in renal damage progression, including hemodynamic alterations, oxidative stress, interstitial inflammation, and tubulointerstitial fibrosis. Key players in the pathogenesis of renal damage, such as the renin-angiotensin-aldosterone system, reactive oxygen species, immune cells, and fibrogenic factors, are discussed in detail. Furthermore, potential therapeutic targets, including renin-angiotensin inhibitors, antioxidants, anti-inflammatory agents, and antifibrotic strategies, are identified based on preclinical and experimental studies. Additionally, emerging therapeutic modalities like mesenchymal stem cells and extracellular vesicles derived from MSCs are explored for their potential in attenuating renal damage and promoting tissue repair. Understanding the pathophysiological mechanisms and identifying potential therapeutic targets is crucial for the development of effective interventions to mitigate renal damage in obstructive uropathies, ultimately improving patient outcomes and quality of life. Further research and clinical trials are needed to translate these promising findings into clinical practice and address the unmet therapeutic needs in this patient populatio