Genomic Stability of the Cells during hiPSC Reprogramming and Endothelial Differentiation
| Autoři | |
|---|---|
| Rok publikování | 2016 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Studies of endothelial biology at genetic and molecular level are limited by the availability of relevant endothelial cells (ECs). Human induced pluripotent stem cells (hiPSCs) offer a potentially unlimited source of ECs. hiPSC-derived ECs (hiPSC-ECs) provide a robust and reproducible patient-specific model system for (1) tissue engineering, (2) drug development, (3) toxicity screening, and (4) disease modeling. However, one of the main concerns is the maintenance of genomic integrity of the cells throughout the processes of hiPSC reprogramming and differentiation in vitro. In our study we generated hiPSCs from various cell-type sources, including human umbilical vein ECs (HUVECs) and adult vein ECs. After phenotypical and functional characterization, hiPSC lines were differentiated into the hiPSC-ECs using previously published protocol (Orlova et al., 2014). Upon magnetic beads-based purification, the hiPSC-EC population displayed endothelial surface markers and functions consistent with primary ECs. In order to provide with global overview of the genome stability and integrity level of the cells throughout the in vitro manipulation, we focused on occurrence of spontaneous double strand DNA breaks (DSBs), functionality of reparation mechanisms, appearance of sub-chromosomal aberrations and differentiation potential of hiPSC sample set. The cells in each stage of the process (source cells – hiPSCs – hiPSC-ECs) were subjected to genomic analysis. Levels of DSBs were assessed using fluorescence microscope in 3D and sub-chromosomal aberrations were analyzed with comparative genomic hybridization (CGH) microarrays. Our study aims to contribute to efforts to eliminate or minimize chromosomal aberrations before hiPSCs will fully realize their scientific and therapeutic potential. |
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