Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells
Pluripotent stem cells (PSC) possess unlimited proliferation, self-renewal, and a differentiation capacity spanning all germ layers. Appropriate culture conditions are important for the maintenance of self-renewal, pluripotency, proliferation, differentiation, and epigenetic states. Oxygen concentra...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-05-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/10/5854 |
| _version_ | 1797499087128363008 |
|---|---|
| author | Fatma Dogan Rakad M. Kh Aljumaily Mark Kitchen Nicholas R. Forsyth |
| author_facet | Fatma Dogan Rakad M. Kh Aljumaily Mark Kitchen Nicholas R. Forsyth |
| author_sort | Fatma Dogan |
| collection | DOAJ |
| description | Pluripotent stem cells (PSC) possess unlimited proliferation, self-renewal, and a differentiation capacity spanning all germ layers. Appropriate culture conditions are important for the maintenance of self-renewal, pluripotency, proliferation, differentiation, and epigenetic states. Oxygen concentrations vary across different human tissues depending on precise cell location and proximity to vascularisation. The bulk of PSC culture-based research is performed in a physiologically hyperoxic, air oxygen (21% O<sub>2</sub>) environment, with numerous reports now detailing the impact of a physiologic normoxia (physoxia), low oxygen culture in the maintenance of stemness, survival, morphology, proliferation, differentiation potential, and epigenetic profiles. Epigenetic mechanisms affect multiple cellular characteristics including gene expression during development and cell-fate determination in differentiated cells. We hypothesized that epigenetic marks are responsive to a reduced oxygen microenvironment in PSCs and their differentiation progeny. Here, we evaluated the role of physoxia in PSC culture, the regulation of DNA methylation (5mC (5-methylcytosine) and 5hmC (5-hydroxymethylcytosine)), and the expression of regulatory enzyme DNMTs and TETs. Physoxia enhanced the functional profile of PSC including proliferation, metabolic activity, and stemness attributes. PSCs cultured in physoxia revealed the significant downregulation of DNMT3B, DNMT3L, TET1, and TET3 vs. air oxygen, accompanied by significantly reduced 5mC and 5hmC levels. The downregulation of DNMT3B was associated with an increase in its promoter methylation. Coupled with the above, we also noted decreased HIF1A but increased HIF2A expression in physoxia-cultured PSCs versus air oxygen. In conclusion, PSCs display oxygen-sensitive methylation patterns that correlate with the transcriptional and translational regulation of the de novo methylase DNMT3B. |
| first_indexed | 2024-03-10T03:42:25Z |
| format | Article |
| id | doaj.art-5510668da6f44894b61c340f4021dd4a |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:42:25Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5510668da6f44894b61c340f4021dd4a2023-11-23T11:29:34ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-012310585410.3390/ijms23105854Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem CellsFatma Dogan0Rakad M. Kh Aljumaily1Mark Kitchen2Nicholas R. Forsyth3The Guy Hilton Research Laboratories, School of Pharmacy and Bioengineering, Faculty of Medicine and Health Sciences, Keele University, Stoke on Trent ST4 7QB, UKDepartment of Biology, College of Science, University of Baghdad, Baghdad 17635, IraqThe Guy Hilton Research Laboratories, School of Pharmacy and Bioengineering, Faculty of Medicine and Health Sciences, Keele University, Stoke on Trent ST4 7QB, UKThe Guy Hilton Research Laboratories, School of Pharmacy and Bioengineering, Faculty of Medicine and Health Sciences, Keele University, Stoke on Trent ST4 7QB, UKPluripotent stem cells (PSC) possess unlimited proliferation, self-renewal, and a differentiation capacity spanning all germ layers. Appropriate culture conditions are important for the maintenance of self-renewal, pluripotency, proliferation, differentiation, and epigenetic states. Oxygen concentrations vary across different human tissues depending on precise cell location and proximity to vascularisation. The bulk of PSC culture-based research is performed in a physiologically hyperoxic, air oxygen (21% O<sub>2</sub>) environment, with numerous reports now detailing the impact of a physiologic normoxia (physoxia), low oxygen culture in the maintenance of stemness, survival, morphology, proliferation, differentiation potential, and epigenetic profiles. Epigenetic mechanisms affect multiple cellular characteristics including gene expression during development and cell-fate determination in differentiated cells. We hypothesized that epigenetic marks are responsive to a reduced oxygen microenvironment in PSCs and their differentiation progeny. Here, we evaluated the role of physoxia in PSC culture, the regulation of DNA methylation (5mC (5-methylcytosine) and 5hmC (5-hydroxymethylcytosine)), and the expression of regulatory enzyme DNMTs and TETs. Physoxia enhanced the functional profile of PSC including proliferation, metabolic activity, and stemness attributes. PSCs cultured in physoxia revealed the significant downregulation of DNMT3B, DNMT3L, TET1, and TET3 vs. air oxygen, accompanied by significantly reduced 5mC and 5hmC levels. The downregulation of DNMT3B was associated with an increase in its promoter methylation. Coupled with the above, we also noted decreased HIF1A but increased HIF2A expression in physoxia-cultured PSCs versus air oxygen. In conclusion, PSCs display oxygen-sensitive methylation patterns that correlate with the transcriptional and translational regulation of the de novo methylase DNMT3B.https://www.mdpi.com/1422-0067/23/10/5854pluripotent stem cellscharacterisationepigeneticmethylationhydroxymethylationphysiological oxygen |
| spellingShingle | Fatma Dogan Rakad M. Kh Aljumaily Mark Kitchen Nicholas R. Forsyth Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells International Journal of Molecular Sciences pluripotent stem cells characterisation epigenetic methylation hydroxymethylation physiological oxygen |
| title | Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells |
| title_full | Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells |
| title_fullStr | Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells |
| title_full_unstemmed | Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells |
| title_short | Physoxia Influences Global and Gene-Specific Methylation in Pluripotent Stem Cells |
| title_sort | physoxia influences global and gene specific methylation in pluripotent stem cells |
| topic | pluripotent stem cells characterisation epigenetic methylation hydroxymethylation physiological oxygen |
| url | https://www.mdpi.com/1422-0067/23/10/5854 |
| work_keys_str_mv | AT fatmadogan physoxiainfluencesglobalandgenespecificmethylationinpluripotentstemcells AT rakadmkhaljumaily physoxiainfluencesglobalandgenespecificmethylationinpluripotentstemcells AT markkitchen physoxiainfluencesglobalandgenespecificmethylationinpluripotentstemcells AT nicholasrforsyth physoxiainfluencesglobalandgenespecificmethylationinpluripotentstemcells |