RESUMEN
Mesenchymal stem cells (MSC), like macrophages, can be polarized in vitro. In particular, activation of type 4 Toll-like receptor in MSC leads to the appearance of the so-called "proinflammatory" MSC phenotype (MSC1). We showed that secretome (conditioned media) of MSC1 can affect the wound healing processes: promote healing and modulate exudative inflammation and subsequent fibroplastic processes in the damaged area. These effects of secretomes of polarized MSC were superior to those of intact MSC.
Asunto(s)
Células Madre Mesenquimatosas/metabolismo , Receptor Toll-Like 4/metabolismo , Animales , Medios de Cultivo Condicionados/farmacología , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Ratas , Ratas Wistar , Cicatrización de Heridas/efectos de los fármacosRESUMEN
We propose a cell model of the human small intestinal wall based on genetically modified Caco-2 cells that allows visualization and quantitative assessment of activation of NF-κB factor and related intracellular pathway by using fluorescence microscopy. A dose-dependent increase in fluorescence intensity of the obtained cells in response to TNFα exposure in concentrations of 1-100 ng/ml was demonstrated. It was found that this parameter correlates with a decrease in the transepithelial resistance of the cell monolayer in response to TNFα and can be used to assess the toxic effects of substances on epithelial cells of the human small intestine.
Asunto(s)
Intestino Delgado/citología , Células CACO-2 , Células Epiteliales/metabolismo , Humanos , Proteínas Luminiscentes/metabolismo , FN-kappa B/metabolismo , Uniones Estrechas/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OMERO service was used to annotate the cell line HaCaT microscope images by two independent expert groups. The images were obtained in the course of developing tissue-engineered epithelium which consisted of several layers of the keratinocytes. Evaluation of expert opinions was performed by calculation of specificity, sensitivity and accuracy. The best convergence of opinions (91%) was achieved for the confluence of the cell monolayers. Accuracy 70% was observed in determining the extent of cell differentiation after 10 days of incubation. The paper illustrates the usefulness of OMERO service for dynamic cross-validation of quality in the development and standardization of cell preparations.