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The interplay between mesenchymal stem/stromal cells (MSCs) and preservation conditions is critical to maintain the viability and functionality of these cells before administration. We observed that Ringer lactate (RL) maintained high viability of bone marrow-derived MSCs for up to 72 h at room temperature (18°C-22°C), whereas adipose-derived and umbilical cord-derived MSCs showed the highest viability for 72 h at a cold temperature (4°C-8°C). These cells maintained their adherence ability with an improved recovery rate and metabolic profiles (glycolysis and mitochondrial respiration) similar to those of freshly harvested cells. Growth factor and cytokine analyses revealed that the preserved cells released substantial amounts of leukaemia inhibitory factors (LIFs), hepatocyte growth factor (HGF) and vascular endothelial growth factor-A (VEGF-A), as well as multiple cytokines (eg IL-4, IL-6, IL-8, MPC-1 and TNF-α). Our data provide the simplest clinically relevant preservation conditions that maintain the viability, stemness and functionality of MSCs from perinatal and adult tissue sources.
Assuntos
Criopreservação , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Tecido Adiposo/citologia , Biomarcadores , Células da Medula Óssea/citologia , Criopreservação/métodos , Citocinas/metabolismo , Metabolismo Energético , Feminino , Humanos , Masculino , Cordão Umbilical/citologiaRESUMO
Introduction: In an effort of gaining a better understanding of the lens mechanics, ex vivo lenses samples are often used. Yet, ex vivo tissue might undergo important postmortem changes depending on the unavoidable preservation method employed. The purpose of this study was to assess how various storage conditions and the removal of the lens capsule affect the mechanical properties of ex vivo porcine lens samples. Methods: A total of 81 freshly enucleated porcine eyes were obtained and divided into six groups and preserved differently. In the first three groups, the lens within the intact eye was preserved for 24 h by: (i) freezing at -80°C (n = 12), (ii) freezing at -20°C (n = 12), and (iii) refrigeration at +8°C (n = 12). In the remaining groups, the lenses were immediately extracted and treated as follows: (iv) kept intact, no storage (n = 12), (v) decapsulated, no storage (n = 21), and (vi) immersed in Minimum Essential Medium (MEM) at +8°C (n = 12) for 24 h. Frozen lenses were thawed at room temperature. Each lens was compressed between two glass lamella and subjected, first to a period of relaxation during which the compression force was recorded and second to an oscillating micro-compression while the deformation was recorded with a total of 256 subsequent B-scans via optical coherence tomography. The corresponding axial strain was retrieved via phase-sensitive image processing and subsequently used as input for an inverse finite element analysis (iFEA) to retrieve the visco-hyperelastic material properties of the lenses. Results: After freezing at temperatures of -80°C and -20°C, the cortical strains increased by 14% (p = 0.01) and 34% (p < 0.001), and the nuclear strains decreased by 17% (p = 0.014) and 36% (p < 0.001), compared to the lenses tested immediately after postmortem, respectively. According to iFEA, this resulted from an increased ratio of the nuclear: cortical E-modulus (4.06 and 7.06) in -80°C and -20°C frozen lenses compared to fresh lenses (3.3). Decapsulation had the largest effect on the material constant C10, showing an increase both in the nucleus and cortex. Preservation of the intact eye in the refrigerator induced the least mechanical alterations in the lens, compared to the intact fresh condition. Discussion: Combining iFEA with optical coherence elastography allowed us to identify important changes in the lens mechanics induced after different preserving ex vivo methods.
RESUMO
The Youjiang Basin is the main area of the Middle Devonian shale gas resources in southern China, with complex geological conditions, diverse stratigraphic structures, low level of exploration and limited hydrocarbon geological data. In this study, a method is proposed for evaluating the regional shale gas preservation conditions, by which it is easy to obtain information (using available data from regional geological maps) and to process data (processed by computer software). This method was applied to the evaluation of the preservation conditions of the Middle Devonian shale gas in the Youjiang Basin. Information extracted from geologic maps includes exposed stratum, magmatic rock distribution, stratigraphic occurrence and surface fracture distribution. Evaluation criteria for three indexes (stratigraphy index, dip angle index and fracture index) were established to classify the preservation conditions into five types from good to bad. Based on the calculated values of comprehensive index (CI), the comprehensive evaluation each area: 11621.23 km2 for CI of 0.7-1.0, 37162.67 km2 for CI of 0.5-0.7, 57784.43 km2 for CI of 0.3-0.5, 69303.77 km2 for CI of 0.1-0.3, and 69303.77 km2 for CI of 0.0-0.1, accounting for 3.19%, 10.19%, 15.84%, 18.99% and 51.80% of the whole area, respectively. These results are highly consistent with the actual exploration discoveries, showing that in the northern and central parts of the Guizhong Basin, the southern part of the Xidamingshan Uplift, and the northern part of the Qiannan Basin, there are large areas with good shale gas preservation conditions.
RESUMO
Since the development of shale gas in the Wufeng-Longmaxi Formation in the Sichuan Basin, China's shale gas production and reserves have increased rapidly. The southeastern margin of the Sichuan Basin is located in a normally pressured transition zone, where single well gas production varies greatly under complex geological structures. In order to reveal the shale gas enrichment mechanism and favorable shale gas regions, shale gas samples from production wells were collected from different structures, with the formation pressure coefficient ranging between 0.98 and 1.35. The gas components and carbon isotope characteristics of normally pressured shale gas were investigated. The carbon isotope characteristics of the Wufeng-Longmaxi shale gas from the basin scale was mainly controlled using thermal maturity; as the thermal maturity increased, heavier carbon isotopes were found, in addition to drier shale gas. For normally pressured shale gas, the composition of δ13C1 and δ13C2 becomes heavier, and the dryness coefficient decreases with the decreasing pressure coefficient; this is not consistent with the results from thermal evolution. By comparing possible influencing factors, it is evident that the change in geological structure destroys the original shale gas reservoir, which leads to the escape of some gases, and it may be the main factor that contributes to the gas geochemical characteristics of the normally pressured shale gas. Compared with the geological parameters of the shale samples, such as mineral composition, organic abundance, organic pore distribution, and gas content, the carbon isotope characteristics of normally pressured shale gas show a higher efficiency, thus indicating favorable sweet spot evaluations for shale gas in the studied areas.