ISOLATION AND DIFFERENTIATION OF STEM CELLS: SEARCHING FOR NEW MARKERS

Abstract MSCs are the most studied adult stem cells, as they can be isolated from almost any tissue, they show a good self-renewal capacity in vitro and they also possess good \u201cplasticity\u201d(jiang nature). Oddly, MSCs are identified and defined by a combination of markers that are not distinctive, as they are shared by other cells including fibroblasts. Therefore, pure populations of MSCs cannot be isolated, as they are always contaminated by other adult cells that often do not possess stem cell plasticity. Thus, it would be very desirable to discover novel cell surface markers that would allow to discriminate MSCs from other cells. Moreover, new ways to control and promote cell differentiation are needed. Thus, it becomes crucial to search for new key molecules that can be modulated to increase MSCs differentiation toward the desired tissue or inhibit the differentiation when cells have to remain in an undifferentiated state. In this direction, we focused our attention on SLs, a family of lipids found in the outer leaflet of the plasma membrane and involved in many cell signaling pathways. Therefore, main aims of this work were: 1. to investigate the possible use of SLs as new surface markers for the identification, characterization and possibly isolation of human bone marrow MSCs; 2. to investigate the involvement of SLs in the preservation of the undifferentiated state of MSCs during in vitro culturing 3. to assess the possible role of SLs in the differentiation processes of MSCs upon opportune stimuli

Life Cycle Assessment (LCA): New poplar clones allow an environmentally sustainable cultivation

In Italy 72 poplar clones ( Populus spp.) are registered for commercialization. They were selected for fast growth, stem shape and disease resistance. The new selections (named MSA) includes genotypes with very high resistance to all the main diseases and to one insect, Phloeomizus passerinii (Sign.). Fast growth and disease resistance allow to produce wood with low environmental and economic costs; for this reason in some Italian Regions the introduction of a percentage of these clones in poplar stand is mandatory to obtain funding for their establishment (Rural Development Plan). To better understand the environmental advantages deriving from the use of these clones, in comparison with the old genotypes (particularly ‘I-214’), a ‘Life Cycle Assessment’ approach was applied considering as impact indicator the CO 2 equivalent emissions; from stoolbed to commercial stand, primary data were collected from an Italian experience. Firstly with the Inventory Analysis all the raw material, energy, wastes and emissions related were collected for each cultivation phase. The Analysis showed a reduction of 9% of CO 2 eq. ha -1 emitted, growing MSA instead of ‘I-214’. Considering the emissions per volume of wood, ‘I-214’ requests 47.5 kg CO 2 eq. per m 3 , compared with MSA that request 36.6 kg CO 2 eq. per m 3

Bioethanol from poplar clone Imola: an environmentally viable alternative to fossil fuel?

BACKGROUND: Environmental issues, e.g. climate change, fossil resource depletion have triggered ambitious national/regional policies to develop biofuel and bioenergy roles within the overall energy portfolio to achieve decarbonising the global economy and increase energy security. With the 10 % binding target for the transport sector, the Renewable Energy Directive confirms the EU’s commitment to renewable transport fuels especially advanced biofuels. Imola is an elite poplar clone crossed from Populus deltoides Bartr. and Populus nigra L. by Research Units for Intensive Wood Production, Agriculture Research Council in Italy. This study examines its suitability for plantation cultivation under short or very short rotation coppice regimes as a potential lignocellulosic feedstock for the production of ethanol as a transport biofuel. A life cycle assessment (LCA) approach was used to model the cradle-to-gate environmental profile of Imola-derived biofuel benchmarked against conventional fossil gasoline. Specific attention was given to analysing the agroecosystem fluxes of carbon and nitrogen occurring in the cultivation of the Imola biomass in the biofuel life cycle using a process-oriented biogeochemistry model (DeNitrification-DeComposition) specifically modified for application to 2G perennial bioenergy crops and carbon and nitrogen cycling. RESULTS: Our results demonstrate that carbon and nitrogen cycling in perennial crop–soil ecosystems such as this example can be expected to have significant effects on the overall environmental profiles of 2G biofuels. In particular, soil carbon accumulation in perennial biomass plantations is likely to be a significant component in the overall greenhouse gas balance of future biofuel and other biorefinery products and warrants ongoing research and data collection for LCA models. We conclude that bioethanol produced from Imola represents a promising alternative transport fuel offering some savings ranging from 35 to 100 % over petrol in global warming potential, ozone depletion and photochemical oxidation impact categories. CONCLUSIONS: Via comparative analyses for Imola-derived bioethanol across potential supply chains, we highlight priority issues for potential improvement in 2G biofuel profiling. Advanced clones of poplar such as Imola for 2G biofuel production in Italy as modelled here show potential to deliver an environmentally sustainable lignocellulosic biorefinery industry and accelerate advanced biofuel penetration in the transport sector. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0318-8) contains supplementary material, which is available to authorized users

Biomass production and energy balance of herbaceous and woody crops on marginal soils in the Po Valley

A wealth of data and information on the cultivation of perennial biomass crops has been collected, but direct comparisons between herbaceous and woody crops are rare. The main objective of this research was to compare the biomass yield, the energy balance and the biomass quality of six perennial bioenergy crops: Populus spp., Robinia pseudoacacia, Salix spp., Arundo donax, Miscanthus 7 giganteus, and Panicum virgatum, grown in two marginal environments. For giant reed and switchgrass, two levels of nitrogen fertilization were applied annually (0-100 kg ha-1). Nitrogen fertilization did not affect biomass or energy production of giant reed; thus, it significantly reduced the energy return on investment (EROI) (from 73 to 27). In switchgrass, nitrogen fertilization significantly increased biomass production and the capacity of this crop to respond to water availability, making it a favorable option when only biomass production is a target. Net energy gain (NEG) was higher for herbaceous crops than for woody crops. In Casale, EROI calculated for poplar and willow (7, on average) was significantly lower than that of the other crops (14, on average). In Gariga, the highest EROI was calculated for miscanthus (98), followed by nonfertilized giant reed and switchgrass (82 and 73, respectively). Growing degree days10 during the cropping season had no effect on biomass production in any of the studied species, although water availability from May to August was a major factor affecting biomass yield in herbaceous crops. Overall, herbaceous crops had the highest ranking for bioenergy production due to their high biomass yield, high net energy gain (NEG), and biomass quality that renders them suitable to both biochemical and thermochemical conversion. Miscanthus in particular had the highest EROI in both locations (16 and 98, in Casale and Gariga), while giant reed had the highest NEG on the silty-loam soil of Gariga

Endothelial function and serum concentration of toxic metals in frequent consumers of fish

BACKGROUND:Endothelial dysfunction is involved in the pathogenesis of atherosclerosis. Consumption of fish is associated with reduced cardiovascular risk, but there is paucity of data concerning its effect on endothelial function. Furthermore, investigation of the effects of fish consumption on health must take into account the ingestion of contaminants, including transition metals and some metalloids, which may have unfavorable effects on health, including those on the cardiovascular system. We investigated the association between fish consumption, endothelial function (flow mediated dilation of the brachial artery), and serum concentration of some toxic metals in apparently healthy people. METHODS:Twenty-nine high fish consumers (at least 3 portions a week) were compared with 25 low fish consumers (less than 1 portion a week). All participants were free of diabetes, cardiovascular or other systemic diseases. Serum metal (antimonium, arsenic, mercury, lead, cobalt, copper, zinc, selenium, strontium) concentrations were measured in subgroups of 24 high fish consumers and 19 low fish consumers. RESULTS:Both groups exhibited similar habitual dietary patterns, age and anthropometric characteristics. The high fish consumers had higher flow mediated dilation (9.7 ± 1.8 vs. 7.3 ± 1.9%; P<0.001), but also higher serum concentrations of mercury (5.87 ± 2.69 vs. 1.65 ± 1.10 mcg/L; P<0.001) and arsenic (6.04 ± 3.25 vs. 2.30 ± 1.58 mcg/L; P<0.001). The fasting plasma glucose concentrations were significantly correlated with both mercury (r = 0.39; P = 0.01) and arsenic concentrations (r = 0.55; P<0.001). CONCLUSIONS:Habitual consumption of high amounts of fish is associated with better endothelial function despite higher serum concentrations of mercury and arsenic

Chemical Activation of the Hypoxia-Inducible Factor Reversibly Reduces Tendon Stem Cell Proliferation, Inhibits Their Differentiation, and Maintains Cell Undifferentiation

Adult stem cell-based therapeutic approaches for tissue regeneration have been proposed for several years. However, adult stem cells are usually limited in number and difficult to be expanded in vitro, and they usually tend to quickly lose their potency with passages, as they differentiate and become senescent. Culturing stem cells under reduced oxygen tensions (below 21%) has been proposed as a tool to increase cell proliferation, but many studies reported opposite effects. In particular, cell response to hypoxia seems to be very stem cell type specific. Nonetheless, it is clear that a major role in this process is played by the hypoxia inducible factor (HIF), the master regulator of cell response to oxygen deprivation, which affects cell metabolism and differentiation. Herein, we report that a chemical activation of HIF in human tendon stem cells reduces their proliferation and inhibits their differentiation in a reversible and dose-dependent manner. These results support the notion that hypoxia, by activating HIF, plays a crucial role in preserving stem cells in an undifferentiated state in the "hypoxic niches" present in the tissue in which they reside before migrating in more oxygenated areas to heal a damaged tissue

GM1 Ganglioside Promotes Osteogenic Differentiation of Human Tendon Stem Cells

Gangliosides, the sialic acid-conjugated glycosphingolipids present in the lipid rafts, have been recognized as important regulators of cell proliferation, migration, and apoptosis. Due to their peculiar localization in the cell membrane, they modulate the activity of several key cell receptors, and increasing evidence supports their involvement also in stem cell differentiation. In this context, herein we report the role played by the ganglioside GM1 in the osteogenic differentiation of human tendon stem cells ( hTSCs). In particular, we found an increase of GM1 levels during osteogenesis that is instrumental for driving the process. In fact, supplementation of the ganglioside in the medium significantly increased the osteogenic differentiation capability of hTSCs. Mechanistically, we found that GM1 supplementation caused a reduction in the phosphorylation of the platelet-derived growth factor receptor-ss ( PDGFR-ss), which is a known inhibitor of osteogenic commitment. These results were further corroborated by the observation that GM1 supplementation was able to revert the inhibitory effects on osteogenesis when the process was inhibited with exogenous PDGF

Lipogems product treatment increases the proliferation rate of human tendon stem cells without affecting their stemness and differentiation capability

Increasing the success rate of rotator cuff healing remains tremendous challenge. Among many approaches, the possibility of activating resident stem cells in situ, without the need to isolate them from biopsies, could represent valuable therapeutic strategy. Along this line, it has been recently demonstrated that lipoaspirate product, Lipogems, contains and produces growth-factors that may activate resident stem cells. In this study, human tendon stem cells (hTSCs) from the rotator cuff were cocultured in a transwell system with the Lipogems lipoaspirate product and compared to control untreated cells in terms of cell proliferation, morphology, stem cell marker and VEGF expression, and differentiation and migration capabilities. Results showed that the Lipogems product significantly increases the proliferation rate of hTSCs without altering their stemness and differentiation capability. Moreover, treated cells increase the expression of VEGF, which is crucial for the neovascularization of the tissue during the healing process. Overall, this study supports that directly activating hTSCs with the Lipogems lipoaspirate could represent a new practical therapeutic approach. In fact, obtaining a lipoaspirate is easier, safer, and more cost-effective than harvesting cells from tendon or bone marrow biopsies, expanding them in GMP facility and then reinjecting them in the patient