The characteristics of endothelial progenitor cells derived from mononuclear cells of rat bone marrow in different culture conditions

Endothelial progenitor cells (EPCs) derived from bone marrow are known to be heterogeneous. In this study, we tried to find favorable conditions that induce the differentiation of mononuclear cells (MNCs) from bone marrow into EPCs. The differentiation capacity of MNCs from rat bone marrow was investigated in different conditions, such as different media, different induction times and different culture surfaces. The cell morphology and endothelial biomarkers associated with differentiated MNCs were studied. Our results indicated that MNCs cultured in EGM-2MV (Endothelial cell basal medium-2, plus SingleQuots of growth supplements) developed a bursiform shape, a late EPC-like morphology, while MNCs cultured in complete medium (CM, M199 with 10% FBS, 20 ng/mL VEGF and 10 ng/mL bFGF) showed a spindle shape, an early EPC-like morphology. Cells of both morphologies were able to incorporate DiI-ac-LDL and bind lectin in vitro. MNCs cultured in EGM-2MV exhibited a higher proliferation rate and higher eNOS expression than MNCs cultured in CM. MNCs cultured in EGM-2MV had the ability to form tubes on Matrigel. Flow cytometry results indicated that CD133 expression was highest at day 12 and that the greatest number of cells positive for both FLK-1 and CD133 appeared at day 20 from cells cultured in dishes without fibronectin coating. In addition, the expression levels of CD133, CD31 and FLK-1/CD133 were not significantly different between cells of different shapes. Our experiments suggest that MNCs from bone marrow can be differentiated into late EP-like cells in EGM-2MV, which have the ability to rapidly proliferate. These MNCs can also be differentiated into early EP-like cells in CM. Additionally, fibronectin may not be necessary for the differentiation of EPCs to mature ECs after three generations. Differentiated MNCs from bone marrow in EGM-2MV have the characteristics of EPCs, although the expression levels of EPC markers were lower than previously reported

Dependência espacial em levantamentos do estoque de carbono em áreas de pastagens de Brachiaria brizantha cv. Marandu Spatial dependence in surveys of carbon storage in grassland areas of Brachiaria brizantha, Marandu grass

Foi conduzido um estudo utilizando análise de semivariogramas para quantificar a autocorrelação espacial dos estoques de carbono (EC) no solo, biomassa da gramínea e das plantas daninhas em três parcelas experimentais de pastagens de Brachiaria brizantha cv. Marandu com níveis baixo, médio e alto de degradação, cultivadas em Neossolo Quartzarênico Órtico. As coletas das plantas e do solo foram realizadas em malha de amostragem regular com distâncias de 5 x 5 m em área de 900 m². Os EC das pastagens foram submetidos às analises de estatística descritiva, ao teste não-paramétrico de Kruskal-Wallis ao nível de 5% de significância, ao estudo geoestatístico e interpolação por krigagem ordinária. A variabilidade espacial do EC foi observada dentro e entre as pastagens de capim-Marandu com níveis baixo, médio e alto de degradação. A pastagem de capim-Marandu com nível baixo de degradação teve menor continuidade espacial, por apresentar menores alcances no EC, na biomassa da gramínea e na biomassa total (gramínea + plantas daninhas), no solo e no sistema solo x pastagem (solo + biomassa total). A grade de 5 x 5 m foi adequada para caracterizar a variabilidade espacial de pastagens de capim-Marandu com níveis de degradação baixo e alto. Área de pastagem de capim-Marandu com grau médio de degradação apresenta coeficientes de variação altos entre os valores EC; o que comprometeu a modelagem espacial que também pode ter ocorrido devido ao baixo número de amostras realizadas (n=36). Assim, pontos de amostragem menores que 5 m podem melhorar a precisão dos ajustes dos semivariogramas.<br>A study was carried using semivariogram analysis to quantify spatial autocorrelation of carbon stock (CS) in soil, grass and weed species biomass in pastures of Brachiaria brizantha, Marandu grass with low, medium and high degradation, and grown an entisol. The sampling of plants and soil were carried out in regular grid with distances of 5x5 m in an area of 900 m². Grassland CS was assessed through descriptive statistics, comparison of averages through the test Kruskal-Wallis at 5% level of significance, geostatistics and ordinary kriging analysis. The spatial variability of CS was observed within and between pastures with low, medium and high degradation. Pastures with low levels of degradation had less spatial continuity due to smaller ranges in CS in grass biomass and total biomass (grass + weed species), in soil carbon and soil versus grass (total biomass + soil). The grid of 5x5 m was adequate to characterize the spatial variability of pasture with low and high levels of degradation. Areas of Marandu grass with average degree of degradation has high coefficients of variation (CV) between the CS values, which negative by affected the spatial modeling. High CV may also be due to the low number of samples taken (n = 36). Sampling points in grid lower than 5 m can improve the accuracy of the adjustment of semivariograms