No abstract

Expression of major histocompatibility antigens class-2 (MHC-II) under non-inflammatory conditions is not usually associated with the nervous system. Comparative analysis of immunogenicity of human embryonic/fetal brain-derived neural stem cells (hNSCs) and human mesenchymal stem cells with neurogenic potential from umbilical cord (UC-MSCs) and paediatric adipose tissue (ADSCs), while highlighting differences in their immunogenicity, led us to discover subsets of neural cells co-expressing the neural marker SOX2 and MHC-II antigen in vivo during human CNS development. MHC-II proteins in hNSCs are functional, and differently regulated upon differentiation along different lineages. Mimicking an inflammatory response using the inflammatory cytokine IFNγ induced MHC-II up-regulation in both astrocytes and hNSCs, but not in UC-MSCs and ADSCs, either undifferentiated or differentiated, though IFNγ receptor expression was comparable. Together, hypoimmunogenicity of both UC-MSCs and ADSCs supports their suitability for allogeneic therapy, while significant immunogenicity of hNSCs and their progeny may at least in part underlie negative effects reported in some patients following embryonic neural cell grafts. Crucially, we show for the first time that MHC-II expression in developing human brains is not restricted to microglia as previously suggested, but is present in discrete subsets of neural progenitors and appears to be regulated independently of inflammatory stimuli.

Human somatic stem cells with neural differentiation potential can be valuable for developing cell-based therapies, including treatment of birth-related defects, while avoiding issues associated with cell reprogramming. Precisely defining the "identity" and differentiation potential of somatic stem cells from different sources, has proven difficult, given differences in sets of specific markers, protocols used and lack of side-by-side characterization of these cells in different studies. Therefore, we set to compare expression of mesenchymal and neural markers in human umbilical cord-derived mesenchymal stem cells (UC-MSCs), pediatric adipose-derived stem cells (p-ADSCs) in parallel with human neural stem cells (NSCs). We show that UC-MSCs at a basal level express mesenchymal and so-called "neural" markers, similar to that we previously reported for the p-ADSCs. All somatic stem cell populations studied, independently from tissue and patient of origin, displayed a remarkably similar expression of surface markers, with the main difference being the restricted expression of CD133 and CD34 to NSCs. Expression of certain surface and neural markers was affected by the expansion medium used. As predicted, UC-MSCs and p-ADSCs demonstrated tri-mesenchymal lineage differentiation potential, though p-ADSCs display superior chondrogenic differentiation capability. UC-MSCs and p-ADSCs responded also to neurogenic induction by up-regulating neuronal markers, but crucially they appeared morphologically immature when compared with differentiated NSCs. This highlights the need for further investigation into the use of these cells for neural therapies. Crucially, this study demonstrates the lack of simple means to distinguish between different cell types and the effect of culture conditions on their phenotype, and indicates that a more extensive set of markers should be used for somatic stem cell characterization, especially when developing therapeutic approaches.

Diamond is an attractive material for supporting the attachment and development of cells as it can show exceptional biocompatibility. When boron is used as a dopant within diamond it becomes a p-type semiconductor, and at high concentrations the diamond becomes quasi-metallic, offering the prospect of a direct electrical device-cell interfacing system.

This article reviews the development of artificial bone substitutes from their older single-phase forms to novel multi-phase composites, mimicking the composition and architecture of natural bone tissue. The new generation of bone implants should be bioactive, i.e. they should induce the desired cellular responses, leading to integration of the material into the natural tissue and stimulating self-healing processes. Therefore, the first part of the review explains the common principles of the cellmaterial interaction and summarizes the strategies how to improve the biocompatibility and bioactivity of the materials by modifying the physico-chemical properties of the material surface, such as surface chemistry, wettability, electrical charge, rigidity, microroughness and especially nanoroughness. The latter has been shown to stimulate preferentially the growth of osteoblasts in comparison with other competitive cell types, such as fibroblasts, which could prevent fibrous tissue formation upon implantation. The second more specialized part of the review deals with materials suitable for bone contact and substitution, particularly novel polymer-based composites reinforced with fibres or inorganic particles and containing bioactive components, such as crystals of hydroxyapatite or other calcium phosphates, synthetic ligands for cell adhesion receptors or growth factors. Moreover, if they are degradable, they can be gradually replaced with a regenerating tissue.

The adult human central nervous system (CNS) has very limited regenerative capability, and injury at the cellular and molecular level cannot be studied in vivo. Modelling neural damage in human systems is crucial to identifying species-specific responses to injury and potentially neurotoxic compounds leading to development of more effective neuroprotective agents. Hence we developed human neural stem cell (hNSC) 3-dimensional (3D) cultures and tested their potential for modelling neural insults, including hypoxic-ischaemic and Ca 2+ -dependent injury. Standard 3D conditions for rodent cells support neuroblastoma lines used as human CNS models, but not hNSCs, but in all cases changes in culture architecture alter gene expression. Importantly, response to damage differs in 2D and 3D cultures and this is not due to reduced drug accessibility. Together, this study highlights the impact of culture cytoarchitecture on hNSC phenotype and damage response, indicating that 3D models may be better predictors of in vivo response to damage and compound toxicity.In light of the practical and ethical limitations of human research, animal models provide fundamental insight into the central nervous system (CNS) development, injury, disease, as well as the possibility of studying and screening putative therapeutic compounds. However, there is an increasing appreciation of the limitations of animal research, which often fails to reliably predict successful human clinical trials. Key examples of this are hypoxia-ischemia either in newborns (1-8 cases per 1000 births in the Western world) or adults (cerebral stroke) and traumatic injuries to the CNS 1-3 . While animal models have significantly furthered understanding of the mechanisms of injury and repair, and various neuroprotective agents have proven effective in animal injury models, this has not translated into effective treatments in humans 4-6 . Such failures to translate animal studies to successful human clinical trials are often attributed to problems with methodological and study design. However, species-specific differences in drug processing, genetic interactions and molecular mechanisms of action can be argued to be fundamental to therapeutic translation. Further studies on the cell ular mechanisms of human CNS injury and repair are therefore needed in order to develop better therapeutic strategies and bridge the gap between animal models and human clinical trials 7,8 .Three dimensional (3D) culture models have been rapidly emerging as a means to study human cells in vitro. While still in many respects rather challenging, because more complex to maintain and analyse, they reduce the strain and artificial responses which cells must undergo in order to adapt to the flat, stiff surfaces of 2D monolayer culture. A 3D architecture provides a more tissue-like environment in which cell-cell and cell-matrix interact in all dimensions and cells have more biologically relevant exposure to diffusible factors 9,10 . Whereas the use of iPSCs (induced pluripotent stem cells) derived f...

Human umbilical cord blood (hUCB) has been proposed to contain not only haematopoietic stem cells, but also a rare pluripotent embryonic-like stem cell (ELSc) population that is negative for hematopoietic markers (Lin−CD45−) and expresses markers typical of pluripotent cells. The aim of this work was to isolate, characterise and expand this ELSc fraction from hUCB, as it may provide a valuable cell source for regenerative medicine applications. We found that we could indeed isolate a Lin−CD45− population of small cells (3–10 µm diameter) with a high nucleus to cytoplasm ratio that expressed the stem cell markers CD34 and CXCR4. However, in contrast to some previous reports, this fraction was not positive for CD133. Furthermore, although these cells expressed transcripts typical of pluripotent cells, such as SOX2, OCT3/4, and NANOG, they were not able to proliferate in any of the culture media known to support stem cell growth that we tested. Further analysis of the Lin−CD45− population by flow cytometry showed the presence of a Lin−CD45−Nestin+ population that were also positive for CD34 (20%) but negative for CXCR4. These data suggest that the Lin−CD45− stem cell fraction present in the cord blood represents a small heterogeneous population with phenotypic characteristics of stem cells, including a Lin−CD45−Nestin+ population not previously described. This study also suggests that heterogeneity within the Lin−CD45− cell fraction is the likely explanation for differences in the hUCB cell populations described by different groups that were isolated using different methods. These populations have been widely called “embryonic-like stem cell” on the basis of their phenotypical similarity to embryonic stem cells. However, the fact they do not seem to be able to self-renew casts some doubt on their identity, and warns against defining them as “embryonic-like stem cell” at this stage.

The synthesis and biological properties of a structurally novel, potent and non-peptidic inhibitor of peptidylarginine deiminase are described. The novel drug-like PAD inhibitor contains a 3,5dihydroimidazol-4-one ring that replaces the acyclic guanidine-binding unit present in arginine residues.This new drug-like PAD inhibitor was effective at 100 nM or below and could have relevance to diseases in which PAD expression is up-regulated, including rheumatoid arthritis, cancer, multiple sclerosis, and neural injury.