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A diabetic milieu promotes OCT4 and NANOG production in human visceral-derived adipose stem cells
P Dentelli1, C Barale, G Togliatto
1Department of Medical Sciences, University of Torino, Corso Dogliotti 14, 10126 Torino, Italy.
Aims/Hypothesis:
Successful outcomes have been obtained by exploiting adipose-derived stem cells (ASCs) in regenerative medicine. NADPH oxidase (NOX)-generated reactive oxygen species (ROS) are known to control stem cell self-renewal. Several high glucose (HG)-mediated effects depend on NOX-generated ROS. In this study, we investigated whether, and how mechanistically, HG concentrations control ASC fate in patients with diabetes.
Methods:
ASCs from the visceral adipose tissue of non-diabetic (N-ASCs) and diabetic participants (D-ASCs), identified by surface markers, were counted and evaluated for ROS generation and stem cell properties. Their ability to release soluble factors was assessed by BioPlex analysis. To reproduce an in vitro diabetic glucose milieu, N-ASCs were cultured in HG (25 mmol/l) or normal glucose (NG) concentration (5 mmol/l), as control. ASC pluripotency was assessed by in vitro study. The p47(phox) NOX subunit, AKT and octamer-binding transcription factor 4 (OCT4; also known as POU5F1) were knocked down by small-interfering RNA technology. Stem-cell features were evaluated by sphere cluster formation.
Results:
The ASC number was higher in diabetic patients than in non-diabetic controls. Production of OCT4 and NANOG, stem-cell-specific transcription factors, was upregulated in D-ASCs compared with N-ASCs. Moreover, we found that ROS production and AKT activation drove D-ASC, but not N-ASC, secretion. When N-ASCs were cultured in vitro in the presence of HG, they also expressed OCT4/NANOG and formed spheres. By knock-down of the p47(phox) NOX subunit, AKT and OCT4 we demonstrated that NOX-generated ROS and their downstream signals are crucial for HG-mediated ASC de-differentiation and proinflammatory cytokine production.
Conclusions/Interpretation:
We herein provide a rationale for exploiting D-ASCs in regenerative medicine and/or exploiting HG preconditioning to increase ASCs ex vivo.
Insights
Diabetic patients have more adipose-derived stem cells (ASCs) with enhanced self-renewal properties due to high glucose (HG). This suggests HG preconditioning can improve ASCs for regenerative medicine applications.
Area of Science:
- Stem cell biology
- Regenerative medicine
- Metabolic disease research
Background:
- Adipose-derived stem cells (ASCs) are crucial in regenerative medicine.
- NADPH oxidase (NOX)-generated reactive oxygen species (ROS) regulate stem cell self-renewal.
- High glucose (HG) effects are often mediated by NOX-generated ROS.
Purpose of the Study:
- To investigate how high glucose (HG) concentrations influence adipose-derived stem cell (ASC) fate in diabetic patients.
- To elucidate the mechanistic pathways involved in HG-mediated ASC changes.
Main Methods:
- Compared ASCs from non-diabetic (N-ASCs) and diabetic (D-ASCs) individuals.
- Assessed ROS generation, stem cell properties, and secreted factors.
- Cultured N-ASCs in high glucose (HG) conditions in vitro.
- Utilized small-interfering RNA to knock down key genes like p47(phox), AKT, and OCT4.
Main Results:
- Diabetic patients exhibited increased ASC numbers and upregulated stem cell transcription factors (OCT4, NANOG).
- ROS production and AKT activation were linked to D-ASC secretion.
- In vitro HG exposure induced stem cell marker expression and sphere formation in N-ASCs.
- NOX-generated ROS and downstream signaling were critical for HG-induced ASC de-differentiation and cytokine production.
Conclusions:
- High glucose promotes stem cell characteristics in ASCs.
- Diabetic ASCs (D-ASCs) possess properties that could be leveraged for regenerative medicine.
- Ex vivo HG preconditioning may enhance ASCs for therapeutic use.
