Development of perinatal cell spheroids with dual potential for type 1 diabetes therapy: insulin production and immune modulation

Type 1 diabetes mellitus (T1DM) is an autoimmune disorder characterized by the immune system's destruction of pancreatic beta-cells, leading to insulin deficiency. Conventional treatments focus on insulin replacement, which does not address the underlying immune attack against beta-cells. Therefore, considerable research has been directed towards stem cell therapy, specifically taking advantage of three-dimensional (3D) cell cultures for enhancing the immunomodulatory and differentiative capacities of stem cells. The final goal of the study is to develop a cellular therapy for T1DM by using perinatal cells to create spheroids that can release insulin and mitigate autoimmune responses. The aim of this study is to create a reliable cellular model for regenerative medicine applications in T1DM. Specifically, it aims to develop 3D co-culture spheroids of amniotic epithelial cells (AECs) and Wharton's jelly mesenchymal stromal cells (WJ-MSCs) to restore insulin production and protect insulin producing cells from autoimmune destruction. We created co-culture spheroids of AECs and WJ-MSCs in a 1:1 ratio. These spheroids were analyzed for viability, extracellular matrix production, and hypoxic state. The immunomodulatory ability was evaluated by co-culturing with activated PBMCs and T cells. AECs were differentiated into insulin-producing cells, confirmed by immunofluorescence for specific markers, and combined with undifferentiated WJ-MSCs, which have strong immunomodulatory properties. The undifferentiated co-culture spheroids remained stable and viable in long-term culture with consistent extracellular matrix production. Moreover, the perinatal spheroids showed significant immunomodulatory properties, reducing pro-inflammatory cell activation and promoting anti-inflammatory responses. Spheroids with differentiated cells combined with undifferentiated WJ-MSCs successfully formed cohesive structures and showed potential for insulin production. Perinatal spheroids represent a promising dual approach for developing new T1DM treatment. Their ability to modulate the immune system and differentiate into insulin-producing cells may address both the symptoms and underlying causes of T1DM, offering a comprehensive cellular therapy beyond traditional insulin replacement.