Tissue engineering approaches using a single stem cell type have been unsuccessful in achieving regeneration of radiation-damaged salivary glands (SGs). Therefore, we combined adult SG stem cells (SGSCs) and adipose-derived stem cells (ASCs) in optimized spheroids to enhance the regeneration of damaged SGs in mice. SGSC/ASC spheroids survived longer and secreted more vascular endothelial growth factor (VEGF) than did SGSC spheroids. To enhance cell viability and differentiation, SGSC/ASC spheroids were stimulated with the Wnt/β-catenin signaling activator CHIR99021 and retinoic acid inhibitor BMS431 to maintain stemness. FGF7/10 induced the differentiation of multiple stimulated SGSC/ASC spheroids. SGSC/ASC spheroids exhibited improved viability, calcium channel function, differentiation, and VEGF secretion after multiple, sequential stimulation. The 21-day survival rate of multiply stimulated SGSC/ASC spheroids was two-fold that of non-treated SGSC/ASC spheroids. Optimized SGSC/ASC spheroids enhanced the in vivo regeneration and functional recovery of SG tissue in mice by promoting neovascularization. Genes related to VEGF, such as Vegfa and Vegfb, were highly expressed 2∼6 times for in vitro and 1.5 times for in vivo experiment in the multiply stimulated SGSC/ASC spheroids. Therefore, SGSC/ASC spheroids that have undergone multiple stimulation may be useful for healing radiation-damaged SGs.

使用单一干细胞类型的组织工程方法未能成功实现辐射损伤的唾液腺 (SG) 的再生。因此，我们将成体 SG 干细胞 (SGSC) 和脂肪干细胞 (ASC) 组合在优化的球体中，以增强小鼠受损 SG 的再生。SGSC/ASC 球体比 SGSC 球体存活时间更长，分泌更多的血管内皮生长因子 (VEGF)。为了增强细胞活力和分化，用 Wnt/β-连环蛋白信号激活剂 CHIR99021 和视黄酸抑制剂 BMS431 刺激 SGSC/ASC 球体以维持干性。FGF7/10 诱导了多个受刺激的 SGSC/ASC 球体的分化。SGSC/ASC 球体在多次连续刺激后表现出改善的活力、钙通道功能、分化和 VEGF 分泌。多重刺激的 SGSC/ASC 球体的 21 天存活率是未经处理的 SGSC/ASC 球体的两倍。优化的 SGSC/ASC 球体增强了通过促进新生血管形成小鼠体内SG 组织的再生和功能恢复。在多重刺激的 SGSC/ASC 球体中，与 VEGF 相关的基因，如Vegfa和Vegfb，在体外实验中高表达 2∼6 倍，在体内实验中高表达 1.5 倍。因此，经过多次刺激的 SGSC/ASC 球体可能有助于治愈辐射损伤的 SG。