Impacto do soro de mulheres com e sem obesidade sobre o metabolismo, estresse celular e diferenciação de Células-Tronco Neurais humanas

Abstract

Maternal obesity is associated with an increased risk of gestational complications and neurodevelopmental disorders in the offspring. Given the limitations of animal models, Neural Stem Cells (NSCs) derived from human Embryonic Stem Cells (hESCs) have emerged as relevant systems for studying neural development and cellular responses to altered environments. This study aimed to analyze the differentiation of hESCs into NSCs, assess the expression of neural markers throughout this process, and investigate the effects of conditioning with serum from eutrophic and obese women on viability, proliferation, oxidative stress, and neural differentiation. A metabolomic analysis of the sera was also performed. NSC generation was achieved through a two-step protocol, involving neural induction in 3D culture followed by expansion in 2D. Differentiation was confirmed by RT-qPCR and immunocytochemistry, demonstrating the expression of pluripotency and neural markers at distinct stages. Progressive neural differentiation was observed, indicating phenotypic maturation over time in culture. Following validation, NSCs were exposed to human serum and compared to a control group maintained in standard medium. Metabolomic profiling revealed marked alterations in the sera from obese women, reflecting disrupted lipid metabolism, increased oxidative stress, and metabolic imbalance. Exposure to human serum led to reduced cell viability and significant changes in the NSC protein expression profile, including decreased levels of proliferation and neural progenitor markers (KI67, PAX6, SOX1), alongside increased NEUN and P53 expression, suggesting early induction of differentiation and signs of cellular stress. Cells treated with serum from obese women exhibited increased metabolic activity at 48 and 72 hours, likely reflecting greater availability of energy substrates, such as fatty acids. However, this profile was also associated with elevated oxidative stress. Collectively, the findings indicate that serum from obese women exerts deleterious effects on the physiology of human NSCs, particularly by disrupting redox homeostasis. These results highlight the potential impact of obesity on the fetal microenvironment and its influence on neurodevelopment, underscoring the value of human cellular models in investigating systemic metabolic effects on neural biology.