Migration, fate and in vivo imaging of adult stem cells in the CNS

Adult stem cells have been intensively studied for their potential use in cell therapies for neurodegenerative diseases, ischemia and traumatic injuries. One of the most promising cell sources for autologous cell transplantation is bone marrow, containing a heterogenous cell population that can be roughly divided into hematopoietic stem and progenitor cells and mesenchymal stem cells (MSCs). MSCs are multipotent progenitor cells that, in the case of severe tissue ischemia or damage, can be attracted to the lesion site, where they can secrete bioactive molecules, either naturally or through genetic engineering. They can also serve as vehicles for delivering therapeutic agents. Mobilized from the marrow, sorted or expanded in culture, MSCs can be delivered to the damaged site by direct or systemic application. In addition, MSCs can be labeled with superparamagnetic nanoparticles that allow in vivo cell imaging. Magnetic resonance imaging (MRI) is thus a suitable method for in vivo cell tracking of transplanted cells in the host organism. This review will focus on cell labeling for MRI and the use of MSCs in experimental and clinical studies for the treatment of brain and spinal cord injuries.     

Stability of spatially heterogeneous steady-state distributions of oxygen-chemotactic aerobic/anaerobic bacteria

Alternative sufficient conditions are derived that guarantee the stability of spatially heterogenous steady-state distributions of motile aerobic bacterial populations attracted chemotactically by oxygen, motile anaerobic populations repelled by oxygen, and the oxygen concentration itself through a stationary aqueous medium. In particular, it follows from the latter criteria that the heterogeneous steady-state distributions for cylindrical regions with arbitrary cross-sections, uniform depth and mixed Dirichlet-Neumann boundary conditions on the oxygen concentration (appropriate to certain still-water bodies in nature) are stable with respect to arbitrary perturbations in the bacteria cell and the oxygen distributions.     

Isolated adipocytes: an assessment of cell surface changes during their preparation

A method is described, based on the detection of adipocyte-specific cell surface antigens, which allows assessment of the relative surface damage incurred by the cells when they are prepared under a variety of conditions. Using the method it is possible to develop, for any set of reagents, a set of cell isolation conditions (collagenase concentration, time of incubation) which will produce minimally damaged cells which exhibit high levels of specific cell surface immunoreactivity. Under certain conditions a recovery from limited surface damage can be achieved, although, when cells are prepared under more extreme conditions irreversible surface damage occurs. The surface morphology of the cells as revealed by scanning electron microscopy, is also clearly affected by the conditions of cell isolation. The method has been used to define the conditions necessary for the isolation of cells to be used in the study of subtle biochemical responses.     

Regenerative medicine and liver injury: what role for bone marrow derived stem cells?

In recent years, great interest has been aroused by the discovery of the ability of adult stem cells to contribute to regeneration processes and repair of damaged tissues. In particular, bone marrow derived stem cells (BMSCs), the most well known population of multipotent stem cells in adults, have been shown to be able to generate many different committed cellular types. In this review, we systematically organize the numerous hypotheses emerging from the most recent studies, in animal and humans, which evaluated the potentiality of BMSCs to contribute to tissue repair in different types of liver damage. Our aim is to give scientists and clinicians who are interested in regenerative medicine the rational basis for planning future studies on stem cell therapy for liver diseases.     

Ultrastructural Features of Gut Regeneration in Five-Month-Old Pentactulae of the Holothurian Eupentacta fraudatrix

Posterior regeneration of the digestive system after bisection was investigated in the anterior halves of the five-month-old pentactulae of the holothurian Eupentacta fraudatrix using electron microscopy. Three stages of gut restoration were distinguished. The first stage is characterized by degradation of the damaged part of the gut followed by wound healing. Active morphogenetic processes (cell proliferation, dedifferentiation, cell migration, and redifferentiation) are observed at the second stage. During the third (final) stage, the ablated parts differentiate in the posterior portion of the intestine. The cells of the gut remnant tissues were shown to be the cell sources of regeneration. Based on both the data available from the literature and the results of our study, the conclusion was drawn that the mechanisms of gut restoration differ significantly in the pentactulae and adults of E. fraudatrix.     

Stem cells for the replacement of inner ear neurons and hair cells

Stem cells in the nervous system have some capacity to restore damaged tissue. Proliferation of stem cells endows them with self-renewal ability and accounts for in vitro formation of neurospheres, clonally derived colonies of floating cells. However, damage to the nervous system is not readily repaired, suggesting that the stem cells do not provide an easily recruited source of cells for regeneration. The vestibular and auditory organs, despite their limited ability to replace damaged cells, appear to contain cells with stem cell properties. These inner ear stem cells, identified by neurosphere formation and by their expression of markers of inner ear progenitors, can differentiate to hair cells and neurons. Differentiated cells obtained from inner ear stem cells expressed sensory neuron markers and, after co-culture with the organ of Corti, grew processes that extended to hair cells. The neurons expressed synaptic vesicle markers at points of contact with hair cells. Exogenous stem cells have also been used for hair cell and neuron replacement. Embryonic stem cells are one potential source of both hair cells and sensory neurons. Neural progenitors made from embryonic stem cells, transplanted into the inner ear of gerbils that had been de-afferented by treatment with a toxin, differentiated into cells that expressed neuronal markers and grew processes both peripherally into the organ of Corti and centrally. The regrowth of these neurons suggests that it may be possible to replace auditory neurons that have degenerated with neurons that restore auditory function by regenerating connections to hair cells.     

Mesenchymal stromal cells from human perinatal tissues: From biology to cell therapy

Cell-based regenerative medicine is of growing interest in biomedical research. The role of stem cells in this context is under intense scrutiny and may help to define principles of organ regeneration and develop innovative therapeutics for organ failure. Utilizing stem and progenitor cells for organ replacement has been conducted for many years when performing hematopoietic stem cell transplantation. Since the first successful transplantation of umbilical cord blood to treat hematological malignancies, non-hematopoietic stem and progenitor cell populations have recently been identified within umbilical cord blood and other perinatal and fetal tissues. A cell population entitled mesenchymal stromal cells (MSCs) emerged as one of the most intensely studied as it subsumes a variety of capacities: MSCs can differentiate into various subtypes of the mesodermal lineage, they secrete a large array of trophic factors suitable of recruiting endogenous repair processes and they are immunomodulatory.Focusing on perinatal tissues to isolate MSCs, we will discuss some of the challenges associated with these cell types concentrating on concepts of isolation and expansion, the comparison with cells derived from other tissue sources, regarding phenotype and differentiation capacity and finally their therapeutic potential.     

Repair and tolerance of oxidative DNA damage in plants

DNA damage caused by exposure to reactive oxygen species is one of the primary causes of DNA decay in most organisms. In plants, endogenous reactive oxygen species (ROS) are generated not only by respiration and photosynthesis, but also by active responses to certain environmental challenges, such as pathogen attack. Significant extracellular sources of activated oxygen include air pollutants such as ozone and oxidative effects of UV light and low-level ionizing radiation. Plants are well equipped to cope with oxidative damage to cellular macromolecules, including DNA. Oxidative attack on DNA generates both altered bases and damaged sugar residues that undergo fragmentation and lead to strand breaks. Recent advances in the study of DNA repair in higher plants show that they use mechanisms similar to those present in other eukaryotes to remove and/or tolerate oxidized bases and other oxidative DNA lesions. Therefore, plants represent a valuable model system for the study of DNA oxidative repair processes in eukaryotic cells.     

Rapid and robust generation of functional oligodendrocyte progenitor cells from epiblast stem cells

Myelin-related disorders such as multiple sclerosis and leukodystrophies, for which restoration of oligodendrocyte function would be an effective treatment, are poised to benefit greatly from stem cell biology. Progress in myelin repair has been constrained by difficulties in generating pure populations of oligodendrocyte progenitor cells (OPCs) in sufficient quantities. Pluripotent stem cells theoretically provide an unlimited source of OPCs, but current differentiation strategies are poorly reproducible and generate heterogenous populations of cells. Here we provide a platform for the directed differentiation of pluripotent mouse epiblast stem cells (EpiSCs) through defined developmental transitions into a pure population of highly expandable OPCs in 10 d. These OPCs robustly differentiate into myelinating oligodendrocytes in vitro and in vivo. Our results demonstrate that mouse pluripotent stem cells provide a pure population of myelinogenic oligodendrocytes and offer a tractable platform for defining the molecular regulation of oligodendrocyte development and drug screening.     

The fate of damaged mitochondrial DNA in the cell

Mitochondrion is a double membrane organelle that is responsible for cellular respiration and production of most of the ATP in eukaryotic cells. Mitochondrial DNA (mtDNA) is the genetic material carried by mitochondria, which encodes some essential subunits of respiratory complexes independent of nuclear DNA. Normally, mtDNA binds to certain proteins to form a nucleoid that is stable in mitochondria. Nevertheless, a variety of physiological or pathological stresses can cause mtDNA damage, and the accumulation of damaged mtDNA in mitochondria leads to mitochondrial dysfunction, which triggers the occurrence of mitochondrial diseases in vivo. In response to mtDNA damage, cell initiates multiple pathways including mtDNA repair, degradation, clearance and release, to recover mtDNA, and maintain mitochondrial quality and cell homeostasis. In this review, we provide our current understanding of the fate of damaged mtDNA, focus on the pathways and mechanisms of removing damaged mtDNA in the cell.     

Cellular response to oxidative stress: signaling for suicide and survival

Reactive oxygen species (ROS), whether produced endogenously as a consequence of normal cell functions or derived from external sources, pose a constant threat to cells living in an aerobic environment as they can result in severe damage to DNA, protein, and lipids. The importance of oxidative damage to the pathogenesis of many diseases as well as to degenerative processes of aging has becoming increasingly apparent over the past few years. Cells contain a number of antioxidant defenses to minimize fluctuations in ROS, but ROS generation often exceeds the cell's antioxidant capacity, resulting in a condition termed oxidative stress. Host survival depends upon the ability of cells and tissues to adapt to or resist the stress, and repair or remove damaged molecules or cells. Numerous stress response mechanisms have evolved for these purposes, and they are rapidly activated in response to oxidative insults. Some of the pathways are preferentially linked to enhanced survival, while others are more frequently associated with cell death. Still others have been implicated in both extremes depending on the particular circumstances. In this review, we discuss the various signaling pathways known to be activated in response to oxidative stress in mammalian cells, the mechanisms leading to their activation, and their roles in influencing cell survival. These pathways constitute important avenues for therapeutic interventions aimed at limiting oxidative damage or attenuating its sequelae.     

Maintenance and loss of endocytic organelle integrity: mechanisms and implications for antigen cross-presentation

The membranes of endosomes, phagosomes and macropinosomes can become damaged by the physical properties of internalized cargo, by active pathogenic invasion or by cellular processes, including endocytic maturation. Loss of membrane integrity is often deleterious and is, therefore, prevented by mitigation and repair mechanisms. However, it can occasionally be beneficial and actively induced by cells. Here, we summarize the mechanisms by which cells, in particular phagocytes, try to prevent membrane damage and how, when this fails, they repair or destroy damaged endocytic organelles. We also detail how one type of phagocyte, the dendritic cell, can deliberately trigger localized damage to endocytic organelles to allow for major histocompatibility complex class I presentation of exogenous antigens and initiation of CD8⁺ T-cell responses to viruses and tumours. Our review highlights mechanisms for the regulation of endocytic organelle membrane integrity at the intersection of cell biology and immunology that could be co-opted for improving vaccination and intracellular drug delivery.     

Polo-like kinases and Chk2 at the interface of DNA damage checkpoint pathways and mitotic regulation

The cell cycle controls processes of DNA replication and segregation of replicated DNA into two daughter cells. These processes are coordinated by multiple signaling pathways, which employ many protein kinases. The members of the family of Polo-like protein kinases are among these key cell cycle regulators. In response to DNA damage and inhibited DNA replication, DNA structure checkpoints delay cell cycle progression to provide cells with time for repair of damaged DNA and protect it from more severe damage. These effects are achieved by affecting key players of the basic cell cycle regulation of the cells with damaged DNA. This review is focused on the interplay between Chk2, a bona fide checkpoint protein kinase, and Polo-like kinases.     

A ‘fragile cell’ sub‐population revealed during cytometric assessment of Saccharomyces cerevisiae viability in lipid‐limited alcoholic fermentation

Aims: To show that in anaerobic fermentation with limiting lipid nutrients, cell preparation impacts the viability assessment of yeast cells, and to identify the factors involved. Methods and Results: Saccharomyces cerevisiae viability was determined using propidium iodide staining and the flow cytometry. Analyses identified intact cells, dead cells and, under certain conditions, the presence of a third subpopulation of apparently damaged cells. This intermediate population could account for up to 40% of the entire cell population. We describe, analyse and discuss the effects of different solutions for cell resuspension on the respective proportion of these three populations, in particular that of the intermediate population. We show that this intermediate cell population forms in the absence of Ca²⁺/Mg²⁺. Conclusions: Cell preparation significantly impacts population viability assessment by FCM. The intermediate population, revealed under certain conditions, could be renamed as ‘fragile cells’. For these cells, Ca²⁺ and Mg²⁺ reduce cell membrane permeability to PI. Significance and Impact of the Study: This is the first study that analyses and discusses the factors influencing the formation of an intermediate population when studying viability in yeast alcoholic fermentation. With a wider application in biological research, this study provides important support to the relatively new questioning of propidium iodide staining as a universal cell death indicator.     

Restoration of ailing wetlands

It is widely held that humankind's destructive tendencies when exploiting natural resources leads to irreparable harm to the environment. Yet, this thinking runs counter to evidence that many ecological systems damaged by severe natural environmental disturbances (e.g., hurricanes) can restore themselves via processes of natural recovery. The emerging field of restoration ecology is capitalizing on the natural restorative tendencies of ecological systems to build a science of repairing the harm inflicted by humans on natural environment. Evidence for this, for example, comes from a new meta-analysis of 124 studies that synthesizes recovery of impacted wetlands worldwide. While it may take up to two human generations to see full recovery, there is promise, given human will, to restore many damaged wetlands worldwide.     

Age-related loss of skeletal muscle function; impairment of gene expression

Mechano Growth Factor (MGF) is derived from the insulin-like growth factor (IGF-I) but its sequence differs from the systemic IGF-I produced by the liver. MGF is expressed by mechanically overloaded muscle and is involved in tissue repair and adaptation. It is expressed as a pulse following muscle damage and involved in the activation of muscle satellite (stem) cells. These donate nuclei to the muscle fibers that are required for repair and for the hypertrophy processes which may have similar regulatory mechanisms. Muscles in the elderly are unable to upregulate MGF in response to exercise. This is also true in certain diseases and this helps to explain muscle loss in those conditions. There is evidence that MGF is a local tissue repair factor as well as a growth factor and that it has an important role in damage limitation and inducing repair in other post-mitotic tissues. As there is no cell replacement in these tissues there has to be an effective local cellular repair mechanism. With advancing years this seems to become deficient and there is an increased chance that the damaged cells will undergo cell death leading to progressive loss of tissue function.