This research is performed primarily by our collaborators from the Faculty of Medicine with informatics support by CBIA.
Human pluripotent stem cells represent an accessible cell source for novel cell-based clinical research and therapies. Realizing the potential of human induced pluripotent stem cells (hiPSCs), it is possible to produce almost any desired cell type from any patient’s cells. In contrast to embryonic cells, hiPSCs are easier to obtain and do not generate ethical controversy.
We aim to reprogram various human cell types into hiPSCs, for example peripheral blood cells, fibroblasts, or endothelial cells. It is critical that the genomic integrity of the cells remains intact and that the DNA repair systems are fully functional. In our research, we focus on the detection of DNA double-strand breaks (DSBs) by phosphorylated histone H2AX (known as γH2AX) and p53-binding protein 1 (53BP1) in hiPSC lines and compare their status with the differentiated somatic cells (Šimara et al. 2017). For the analysis of the fluorescence microscopy images, we developed an open source software Acquiarium.
hiPSC lines are differentiated in our lab towards several cell types and the process is closely monitored in order to compare the hiPSC-derived cells with their natural counterparts. Epigenomic status of the cells undergoing hematiopoietic differentiation is being assessed by measuring promotor methylation of selected genes (Tesařová et al. 2016; Tesařová et al. 2017). Endothelial differentiation protocols are being improved to increase the yield and quality of endothelial cells (ECs) (Šimara et al. 2018). Currently, ECs are used as in vitro model for studies of vascular development and drug screens or in regenerative medicine. However, ECs from blood vessels or peripheral blood can be obtained in limited numbers and are difficult to expand. We demonstrated that unlimited amount of ECs can be produced from hiPSCs in two weeks according to protocol that is transferable into current good manufacturing practice (cGMP) conditions.
We pay special attention to the cGMP standards. All our hiPSC experiments are conducted in a virus-free and DNA non-integrating setting without feeder cells, which are the main criteria for future clinical applications. In the context of regenerative medicine we focus on projects linked to embryonal stem cell research, medicinal products based on mesenchymal stem cells and gene therapy (CAR technology).