A luciferase-based quick potency assay to predict chondrogenic differentiation.

Chondrogenic differentiation of adipose derived stem cells (ASC) is challenging but highly promising for cartilage repair. Large donor variability of chondrogenic differentiation potential raises the risk for transplantation of cells with reduced efficacy and a low chondrogenic potential. Therefore quick potency assays are required in order to control the potency of the isolated cells before cell transplantation. Current in vitro methods to analyze the differentiation potential are time consuming and thus, a novel enhancer and tissue-specific promoter combination was employed for the detection of chondrogenic differentiation of ASC in a novel quick potency bioassay. Human primary ASC were co-transfected with the Metridia luciferase based collagen type II reporter gene pCMVE_ACDCII-MetLuc together with a Renilla control plasmid and analyzed for their chondrogenic potential. On day 3 after chondrogenic induction, the luciferase activity was induced in all tested donors under three dimensional (3D) culture conditions and in a second approach also under 2D culture conditions. With our newly developed quick potency bioassay we can determine chondrogenic potential already after 3 days of chondrogenic induction and under 2D culture conditions. This will enhance the efficiency of testing cell functionality, which should allow in the future to predict the suitability of cells derived from individual patients for cell therapies, in a very short time and at low costs

Evaluation of saliency tracking as an alternative for health monitoring in PMSM-drives under nonstationary conditions

This paper evaluates the capability of saliency tracking to assess the health condition of permanent magnet synchronous motor (PMSM) drives operating under nonstationary conditions. The evaluated scheme is based on saliency tracking methods, which are associated to the accurate sensorless control of AC drives without zero speed limitations. In this work two representative saliency tracking architectures are evaluated: High Frequency (HF) injection, and PWM transient excitation. Although a monitoring approach based on HF injection was previously reported, a comparative study to evaluate the most representative saliency tracking schemes to assess health condition in drives was still missing. The aim of this work is to fill out this gap by evaluating and comparing two saliency-based monitoring schemes (one based on HF-injection and the other based on PWM transient excitation) to evaluate their performance in the presence of inter-turn winding faults. Simulation and experimental results are presented which confirm that both schemes offer excellent detection capabilities and that are suitable for drives operating under nonstationary conditions including standstill operation. Significant differences are also found for instance, PWM transient excitation offers improved accuracy since the approach is not affected by the inverter nonlinearities and is suitable for full-speed range applications. The main drawback here is complexity and the hardware requirements. Schemes based on HF-injection proved to be very simple and provide comparable results; however a good performance is only guaranteed for the zero-to-medium speed range applications which limit their applicability

Upravljanje vjetroagregatom otporno na me.uzavojske kvarove statorskog namota generatora

Faults of wind turbine generator electromechanical parts are common and very expensive. This paper introduces a generator fault-tolerant control scheme for variable-speed variable-pitch wind turbines that can be applied regardless to the AC generator used. The focus is on generator stator isolation inter-turn fault that can be diagnosed and characterised before triggering the safety device. An extension of the conventional wind turbine control structure is proposed that prevents the fault propagation while power delivery under fault is deteriorated as less as possible compared to healthy machine conditions. Fault-induced and inherent asymmetries of the generator are estimated and respected by the field-oriented control of the generator to eliminate torque oscillations. An approach for asymmetry detection based on an unscented Kalman filter is proposed. Presented fault-tolerant control strategy is developed taking into account its modular implementation and installation in available control systems of existing wind turbines to extend their exploitable life span and increase energy production. Simulation results for the case of a megawatt class wind turbine are presented.Kvarovi elektromehaničkih dijelova generatora vjetroagregata česti su i izrazito skupi. U ovom radu predstavljen je koncept na kvarove otpornog upravljanja generatorom vjetroagregata s promjenjivom brzinom vrtnje i zakretom lopatica primjenjiv bez obzira na tip korištenog izmjeničnog generatora. Naglasak je stavljen na oštećenje izolacije unutar namota jedne faze statora generatora koje se može dijagnosticirati i okarakterizirati prije aktiviranja sustava zaštite. Predložena je nadogradnja postojećeg sustava upravljanja vjetroagregatom koja sprječava širenje kvara i pritom postiže čim manje smanjenje proizvodnje električne energije u odnosu na normalan režim rada. Tako.er je prikazano vektorsko upravljanje generatorom koje uzima u obzir nesimetrije generatora, kako one nastale uslijed djelovanja kvara, tako i one inherentne. U radu je prikazan pristup za otkrivanje nesimetrija generatora zasnovan na nederivacijskom Kalmanovu filtru. Predstavljena strategija upravljanja razvijena je uvažavajući mogućnost modularnog nadovezivanja na postojeće algoritme upravljanja već postavljenih vjetroagregata s ciljem produženja njihova životnog vijeka i povećanja proizvodnje energije. Dani su simulacijski rezultati za slučaj vjetroagregata iz megavatne klase

Perinatal Derivatives: Where Do We Stand? A Roadmap of the Human Placenta and Consensus for Tissue and Cell Nomenclature

Progress in the understanding of the biology of perinatal tissues has contributed to the breakthrough revelation of the therapeutic effects of perinatal derivatives (PnD), namely birth-associated tissues, cells, and secreted factors. The significant knowledge acquired in the past two decades, along with the increasing interest in perinatal derivatives, fuels an urgent need for the precise identification of PnD and the establishment of updated consensus criteria policies for their characterization. The aim of this review is not to go into detail on preclinical or clinical trials, but rather we address specific issues that are relevant for the definition/characterization of perinatal cells, starting from an understanding of the development of the human placenta, its structure, and the different cell populations that can be isolated from the different perinatal tissues. We describe where the cells are located within the placenta and their cell morphology and phenotype. We also propose nomenclature for the cell populations and derivatives discussed herein. This review is a joint effort from the COST SPRINT Action (CA17116), which broadly aims at approaching consensus for different aspects of PnD research, such as providing inputs for future standards for the processing and in vitro characterization and clinical application of PnD.Austrian Science Fund (FWF) DOC 31-B26Medical University GrazUniversita Cattolica del Sacro CuorePRIN 2017 program of Italian Ministry of Research and University (MIUR) 2017RSAFK7Ministry of Health, Italy GR-2018-12366992Slovenian Research Agency - Slovenia P3-0108MRIC UL IP-0510Plan Estatal de Investigacion Cientifica y Tecnica y de InnovacionISCIII Subdireccion General de Evaluacion y Fomento de la InvestigacionMinisterio de Economia y Competitividad, Spain PI16/01642European Union (EU)European Community (EC)German Research Foundation (DFG) GE-2223/2-

Condition monitoring approach for permanent magnet synchronous motor drives based on the INFORM method

This paper proposes a monitoring scheme based on saliency tracking to assess the health condition of PMSM drives operating under non stationary conditions. The evaluated scheme is based on the INFORM methodology, which is associated to the accurate sensorless control of PM drives without zero speed limitation. The result is a monitoring scheme that is able to detect faults that would be very difficult to evaluate under nonstationary conditions. A relevant aspect of the proposed scheme is that it remains valid for full speed range, and can be used for standstill operation. Additionally, the approach is insensitive to the inverter nonlinearities which enhance the detection capabilities further respect to similar topologies. In this work the proposed approach is evaluated numerically and experimentally in the presence of incipient winding faults and inter-turn short circuits in a PM conventional drive. The obtained results show quick response and excellent detection capabilities not only in the detection of faults, but to determine their magnitude which is vital to avoid further degradation

Human Amniotic Epithelial Cell Transplantation Induces Markers of Alternative Macrophage Activation and Reduces Established Hepatic Fibrosis

Chronic hepatic inflammation from multiple etiologies leads to a fibrogenic response that can progress to cirrhosis and liver failure. Transplantation of human amniotic epithelial cells (hAEC) from term delivered placenta has been shown to decrease mild to moderate hepatic fibrosis in a murine model. To model advanced human liver disease and assess the efficacy of hAEC therapy, we transplanted hAEC in mice with advanced hepatic fibrosis. Immunocompetent C57BL/6 mice were administered carbon tetrachloride (CCl4) twice weekly resulting in bridging fibrosis by 12 weeks. hAEC (2×106) were infused via the tail vein at week 8 or weeks 8 and 10 (single and double dose, respectively). Human cells were detected in mouse liver four weeks after transplantation showing hAEC engraftment. CCl4 treated mice receiving single or double hAEC doses showed a significant but similar decrease in liver fibrosis area associated with decreased activation of collagen-producing hepatic stellate cells and decreased hepatic protein levels of the pro-fibrogenic cytokine, transforming growth factor-beta1. CCl4 administration caused hepatic T cell infiltration that decreased significantly following hAEC transplantation. Hepatic macrophages play a crucial role in both fibrogenesis and fibrosis resolution. Mice exposed to CCl4 demonstrated increased numbers of hepatic macrophages compared to normal mice; the number of macrophages decreased significantly in CCl4 treated mice given hAEC. These mice had significantly lower hepatic protein levels of the chemokine monocyte chemoattractant protein-1 than mice given CCl4 alone. Alternatively activated M2 macrophages are associated with fibrosis resolution. CCl4 treated mice given hAEC showed increased expression of genes associated with M2 macrophages including YM-1, IL-10 and CD206. We provide novel data showing that hAEC transplantation induces a wound healing M2 macrophage phenotype associated with reduction of established hepatic fibrosis that justifies further investigation of this potential cell-based therapy for advanced hepatic fibrosis

Changes in Culture Expanded Human Amniotic Epithelial Cells: Implications for Potential Therapeutic Applications

Human amniotic epithelial cells (hAEC) isolated from term placenta have stem cell-like properties, differentiate into tissue specific cells and reduce lung and liver inflammation and fibrosis following transplantation into disease models established in mice. These features together with their low immunogenicity and immunosuppressive properties make hAEC an attractive source of cells for potential therapeutic applications. However, generation of large cell numbers required for therapies through serial expansion in xenobiotic-free media may be a limiting factor. We investigated if hAEC could be expanded in xenobiotic-free media and if expansion altered their differentiation capacity, immunophenotype, immunosuppressive properties and production of immunomodulatory factors. Serial expansion in xenobiotic-free media was limited with cumulative cell numbers and population doubling times significantly lower than controls maintained in fetal calf serum. The epithelial morphology of primary hAEC changed into mesenchymal-stromal like cells by passage 4–5 (P4–P5) with down regulation of epithelial markers CK7, CD49f, EpCAM and E-cadherin and elevation of mesenchymal-stromal markers CD44, CD105, CD146 and vimentin. The P5 hAEC expanded in xenobiotic-free medium differentiated into osteocyte and alveolar epithelium-like cells, but not chondrocyte, hepatocyte, α- and β-pancreatic-like cells. Expression of HLA Class IA, Class II and co-stimulatory molecules CD80, CD86 and CD40 remained unaltered. The P5 hAEC suppressed mitogen stimulated T cell proliferation, but were less suppressive compared with primary hAEC at higher splenocyte ratios. Primary and P5 hAEC did not secrete the immunosuppressive factors IL-10 and HGF, whereas TGF-β1 and HLA-G were reduced and IL-6 elevated in P5 hAEC. These findings suggest that primary and expanded hAEC may be suitable for different cellular therapeutic applications

Different populations and sources of human mesenchymal stem cells (MSC): A comparison of adult and neonatal tissue-derived MSC

The mesenchymal stroma harbors an important population of cells that possess stem cell-like characteristics including self renewal and differentiation capacities and can be derived from a variety of different sources. These multipotent mesenchymal stem cells (MSC) can be found in nearly all tissues and are mostly located in perivascular niches. MSC have migratory abilities and can secrete protective factors and act as a primary matrix for tissue regeneration during inflammation, tissue injuries and certain cancers