Gene expression of human mesenchymal stem cells in 3D cell culture

Two-dimensional (2D) monolayer cell cultures are inevitable for drug development to identify drug candidate molecules because they are extremely potent for screening procedures. They can be used to predict in vivo drug responses for a variety of targets and pathways but they also face disadvantages like a lack of cell-to-cell and cell-to-matrix interaction as well as the loss of tissue-specific architecture. To overcome the drawbacks of 2D systems, three-dimensional (3D) cell culture was designed to provide cells a more physiological environment. In light of the multiple benefits of 3D cell culture it is important to note that this cultivation system is not automated and thus not ideal for high throughput screening (HTS) so far. An automated technique is critical for reproducible and standardized in vitro cultivation and it can greatly speed up preclinical research and total drug development. This procedure also requires a quality control system to monitor results and ensure that it is suitable for customers´ applications. For this purpose, the physiological condition of 2 D and 3D cell cultures will be assessed analyzing gene expression profiles of human mesenchymal stem cells (hMSCs). This will be accomplished by extracting total ribonucleic acid (RNA) from 2D and 3D cells followed by Affymetrix microarrays to get transcription profiles. The resulting data from both cell systems can be compared, and changes in cell behaviour caused by the 3D system can be observed. 3 samples are analyzed, the 2 D standard culture (sample 1) is a reference for the other samples.