The application of physiological oxygen (physoxia) concentrations is becoming increasingly commonplace within a mammalian stem cell culture. Human mesenchymal stem cells (hMSCs) attract widespread interest for clinical application due to their unique immunomodulatory, multi-lineage potential, and regenerative capacities. Descriptions of the impact of physoxia on global DNA methylation patterns in hMSCs and the activity of enzymatic machinery responsible for its regulation remain limited. Human bone marrow-derived mesenchymal stem cells (BM-hMSCs, passage 1) isolated in reduced oxygen conditions displayed an upregulation of SOX2 in reduced oxygen conditions vs. air oxygen (21% O2, AO), while no change was noted for either OCT-4 or NANOG. DNA methylation marks 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) showed decreases in 2% O2 environment (workstation) (2% WKS). DNMT3B (DNA methyltransferase 3B) and TET1 (Ten-eleven translocation enzyme 1) displayed reduced transcription in physoxia. Consistent with transcriptional downregulation, we noted increased promoter methylation levels of DNMT3B in 2% WKS accompanied by reduced DNMT3B and TET1 protein expression. Finally, a decrease in HIF1A (Hypoxia-inducible factor 1A) gene expression in 2% WKS environment correlated with protein levels, while HIF2A was significantly higher in physoxia correlated with protein expression levels vs. AO. Together, these data have demonstrated, for the first time, that global 5mC, 5hmC, and DNMT3B are oxygen-sensitive in hMSCs. Further insights into the appropriate epigenetic regulation within hMSCs may enable increased safety and efficacy development within the therapeutic ambitions.
Background: Adipose derived-mesenchymal stem cells have been used as an alternative to bone marrow cells in this study. Objective: We investigated the in vitro isolation, identification, and differentiation of stem cells into neuron cells, in order to produce neuron cells via cell culture, which would be useful in nerve injury treatment. Method: Mouse adipose mesenchymal stem cells were dissected from the abdominal subcutaneous region. Neural differentiation was induced using β-mercaptoethanol. This study included two different neural stage markers, i.e. nestin and neurofilament light-chain, to detect immature and mature neurons, respectively. Results: The immunocytochemistry results showed that the use of β-mercaptoethanol resulted in
... Show MoreIn the course of generating a library of open-chain epothilones, we discovered a new class of small molecule anticancer agents that has no effect on tubulin but instead kills selected cancer cell lines by harnessing reactive oxygen species in an iron-dependent manner.
Umbilical cord blood (UCB) contains hematopoietic and mesenchymal stem cells(HSCs,MSCs) that have proven useful clinically to reconstitute the hematopoietic system in children and some adults . Fifteen cord blood samples were collected from placenta of newly delivered women in AlKadhemia hospital in Baghdad for normal vaginal delivery. Mono nucleated cells (MNCs) were isolated by using density gradient centrifugation and the MNCs count and viability were determinated by trypan blue.MNCs were cryopreserved using the cryoprotectant solution of 10% concentration of dimethyl sulfoxid (DMSO)using slow cooling and rapid thaw. The aim of the present study
... Show MoreNeural stem cells (NSCs) are progenitor cells which have the ability to self‑renewal and potential for differentiating into neurons, oligodendrocytes, and astrocytes. The in vitro isolation, culturing, identification, cryopreservation were investigated to produce neural stem cells in culture as successful sources for further studies before using it for clinical trials. In this study, mouse bone marrow was the source of neural stem cells. The results of morphological study and immunocytochemistry of isolated cells showed that NSCs can be produced successfully and maintaining their self‑renewal and successfully forming neurosphere for multiple passages. The spheres preserved their morphology in culture and cryopreserved t
... Show MorePluripotent 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% O2) 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
... Show MoreHair is an excellent indicator for abnormal concentration of toxic elements , In this study a random samples from girls hair of 12 cm long were irradiated by a flux of neutrons (4x10^ n/ cm^.s) obtained from an Am-Be neutron source of 5-Ci activitity . The y-ray activity measurements were carried out by using a " 5x5 " well- type Nal (Tl) detector. The study indicates clearly that the maximum concentration of elements was at about 7 cm hair length.