Project description:The development of more complex but reliable systems for compound testing in a pharmaceutical context is a challenging task to date. Three-dimensional (3D), organ mimetic cell culture is aiming to become an alternative to common two-dimensional (2D) cell culture or animal testing in that field. We developed a biocompatible 3D cell culture environment for a hepatocellular carcinoma (HCC) model that enables cellular maintenance in a polycarbonate scaffold structure. Albumin, regarded as a differentiation marker, was elevated in statically 3D cultivated HepG2 cells. Expression of HCC tumor marker alpha-fetoprotein (AFP) was reduced compared to immunofluorescence stainings of 2D cultivated cells. Remarkably, expression of cytokeratin and pathophysiologically relevant beta-1 integrin (ITGB1) was found enhanced in nonperfused 3D cell culture. Changes in gene expression induced by the 3D cultivation environment were investigated using Ingenuity Pathway Analysis (IPA). Our findings revealed involvement of the insulin growth factor (IGF) signaling pathway in upregulation of matrix metalloproteinases (MMP) and ITGB1. The experimental data indicate a more differentiated state in 3D cultivated HepG2 cells than in the respective 2D experiments. Hence, scaffold-supported 3D cultivation of HepG2 cells may lead to a gain of information valuable for both drug testing and cancer research.
Project description:The response of the model cyanobacterium Synechocystis sp. PCC6803 towards light and carbon limitation was systematically probed. To this end, Synechocystis sp. PCC6803 was cultivated in a photo-bioreactor driven in turbidostat-mode. The turbidostat is a continuous cultivation that enabled cells to adapt to a constant environment, leading to a stable and 'optimal' proteome for the respective condition. The major dataset in this project consisted of 5 different 'concentrations' for light and CO2. Changes in the proteome were determined using using liquid chromatography/mass spectrometry and it was found that carbon and light limitation induced gradual but broad responses in gene expression. With decreasing substrate concentration (increasing limitation) a decrease in growth rate and a gradually more severe response in the proteome was visible.
Project description:Korean ginseng (Panax ginseng Meyer) has long been cultivated as an important medicinal plant. Drought results from the moderate water loss, which primarily impairs the growth of ginseng and reduction of yield loss. However, basis of biological clues to understanding the accurate mechanisms related to drought stress in proteome level are still elusive. Therefore, we carried out label-free quantitative proteomic analysis using ginseng roots subjected to drought stress which was grown at less than 10% soil moisture for two weeks, compared with normal ginseng which was grown at 25% soil moisture. The acquired proteins were carried out label-free proteomic analysis using LC-MS/MS. This approach led to the identification of total 2,471 proteins, and out of 195 proteins showed significant modulation. Functional classification revealed that proteins related to secondary metabolites, calcium signaling, and photosynthesis were enriched in control sample (cluster_1), while proteins associated with stress responsive, redox, electron transport, and protein synthesis were mainly dominated in cluster_2 (drought stress condition). Taken together, our results provided an overview of the drought-induced proteomic changes in ginseng root, and it is correlated with physiological changes, contributing to reveal potential marker at proteome level in ginseng.