Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
| Autor: | Odity Mukherjee, Mitradas M. Panicker, Syama Adhikarla, Megha Prakash Bangalore |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Pluripotent Stem Cells Reactive oxygen species metabolism DNA damage Germ layer Genotoxic Stress Ascorbic Acid Biology Antioxidants Article 03 medical and health sciences Genes Reporter Humans Induced pluripotent stem cell Gene Cells Cultured Membrane Potential Mitochondrial Multidisciplinary Glutathione Cell biology Culture Media 030104 developmental biology Reactive Oxygen Species Cell Nucleolus DNA Damage Mutagens |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep42222 |
| Popis: | Culture conditions play an important role in regulating the genomic integrity of Human Pluripotent Stem Cells (HPSCs). We report that HPSCs cultured in Essential 8 (E8) and mTeSR, two widely used media for feeder-free culturing of HPSCs, had many fold higher levels of ROS and higher mitochondrial potential than cells cultured in Knockout Serum Replacement containing media (KSR). HPSCs also exhibited increased levels of 8-hydroxyguanosine, phospho-histone-H2a.X and p53, as well as increased sensitivity to γ-irradiation in these two media. HPSCs in E8 and mTeSR had increased incidence of changes in their DNA sequence, indicating genotoxic stress, in addition to changes in nucleolar morphology and number. Addition of antioxidants to E8 and mTeSR provided only partial rescue. Our results suggest that it is essential to determine cellular ROS levels in addition to currently used criteria i.e. pluripotency markers, differentiation into all three germ layers and normal karyotype through multiple passages, in designing culture media. |
| Databáze: | OpenAIRE |
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