International Journal of Molecular Sciences

Basic fibroblast growth factor (bFGF) and Transforming growth factor-beta (TGF-β) are key regulators of human pluripotent stem cell (hPSC) maintenance, supporting pluripotency and self-renewal. bFGF is particularly critical for sustaining the undifferentiated state and is commonly supplied through feeder-derived conditioned media. Similarly, TGF-β promotes hPSC expansion by modulating signaling pathways and contributing to a supportive stem cell niche. In this study, we investigated how the quality and variability of these growth factors influence hPSC culture performance. To address this, we developed and applied multiple physicochemical characterization methods—including size exclusion and reverse-phase chromatography—to assess growth factor purity and identify impurities across different material sources. Our findings show that certain post-translational modifications in TGF-β (e.g., oxidized variants) did not measurably affect hPSC culture. However, high temperature-dependent instability of bFGF preparations significantly altered hPSC morphology and growth. These findings underscore the need for improved quality control of growth factor components in culture media to ensure consistent hPSC maintenance, thus decreasing variability across experiments. This study highlights the value of correlating analytical physicochemical data with process performance, thereby advancing material understanding, enabling more efficient process development, and facilitating the identification of critical material attributes that affect the quality of cell therapy products.

Sympathetic nervous system (SNS) imbalance is a common pathological basis for cardiovascular diseases, non-alcoholic fatty liver disease, and diabetes. This review focuses on these diseases, analyzing two core mechanisms: excessive sympathetic excitation induced by endoplasmic reticulum stress (ERS) or autophagy dysfunction in key central nuclei (e.g., hypothalamus, rostral ventrolateral medulla); and ERS/autophagy abnormalities in peripheral target organs caused by chronic SNS overactivation. Existing studies confirm that chronic SNS overactivation promotes peripheral metabolic overload via sustained catecholamine release, inducing persistent ERS and disrupting the protective unfolded protein response (UPR)–autophagy network, ultimately leading to cell apoptosis, inflammation, and fibrosis. Notably, central ERS or autophagy dysfunction further perturbs autonomic homeostasis, exacerbating sympathetic overexcitation. This review systematically elaborates on SNS overactivation as a critical bridge mediating UPR–autophagy network dysregulation in central and peripheral tissues, and explores therapeutic prospects of targeting key nodes (e.g., chemical chaperones, specific UPR modulators, nanomedicine), providing a theoretical basis for basic research and clinical translation.

Pancreatic β-cell replacement represents a promising therapeutic avenue for insulin-dependent diabetes, yet clinical translation has been limited by donor scarcity, immune rejection, and incomplete engraftment. Three-dimensional (3D) pancreatic organoids derived from human pluripotent stem cells (hPSCs) or primary tissue offer a scalable and physiologically relevant platform, recapitulating native islet architecture, paracrine interactions, and glucose-responsive insulin secretion. Recent advances in differentiation protocols, vascularization strategies, and immune-protective approaches—including encapsulation and hypoimmunogenic engineering—have enhanced β-cell maturation, survival, and functional performance in vitro and in vivo. Despite these developments, challenges remain in achieving fully mature β-cells, durable graft function, and scalable, reproducible production that is suitable for clinical use. This review highlights the promise of pancreatic organoid engineering, emphasizing strategies to optimize β-cell maturation, vascular integration, and immune protection, and outlines key future directions to advance organoid-based β-cell replacement toward safe, effective, and personalized diabetes therapies.