Health Science Institute, Federal University of Bahia, Salvador, BA, Brazil.
Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
PLoS One. 2022 Dec 19;17(12):e0277863. doi: 10.1371/journal.pone.0277863. eCollection 2022.
Human adipose tissue-derived stem cells (hASC) secretome display various therapeutically relevant effects in regenerative medicine, such as induction of angiogenesis and tissue repair. The benefits of hASC secretome are primarily orchestrated by trophic factors that mediate autocrine and paracrine effects in host cells. However, the composition and the innate characteristics of hASC secretome can be highly variable depending on the culture conditions. Here, we evaluated the combined effect of serum-free media and hypoxia preconditioning on the hASCs secretome composition and biological effects on angiogenesis and wound healing. The hASCs were cultured in serum-free media under normoxic (NCM) or hypoxic (HCM) preconditioning. The proteomic profile showed that pro- and anti-antiangiogenic factors were detected in NCM and HCM secretomes. In vitro studies demonstrated that hASCs secretomes enhanced endothelial proliferation, survival, migration, in vitro tube formation, and in vivo Matrigel plug angiogenesis. In a full-thickness skin-wound mouse model, injection of either NCM or HCM significantly accelerated the wound healing. Finally, hASC secretomes were potent in increasing endothelial density and vascular coverage of resident pericytes expressing NG2 and nestin to the lesion site, potentially contributing to blood vessel maturation. Overall, our data suggest that serum-free media or hypoxic preconditioning enhances the vascular regenerative effects of hASC secretome in a preclinical wound healing model.
人脂肪组织来源的干细胞（hASC）分泌组在再生医学中显示出各种治疗相关的作用，例如诱导血管生成和组织修复。hASC 分泌组的益处主要由营养因子协调，这些营养因子在宿主细胞中介导自分泌和旁分泌作用。然而，hASC 分泌组的组成和固有特性可能会因培养条件的不同而有很大差异。在这里，我们评估了无血清培养基和低氧预处理对 hASC 分泌组组成和对血管生成和伤口愈合的生物学影响的综合作用。将 hASC 在常氧（NCM）或低氧（HCM）预处理下的无血清培养基中培养。蛋白质组学分析表明，NCM 和 HCM 分泌组中存在促血管生成和抗血管生成因子。体外研究表明，hASC 分泌组增强了内皮细胞的增殖、存活、迁移、体外管形成和体内 Matrigel plugs 血管生成。在全层皮肤伤口小鼠模型中，注射 NCM 或 HCM 均可显著加速伤口愈合。最后，hASC 分泌组能够显著增加表达 NG2 和巢蛋白的驻留周细胞的内皮密度和血管覆盖率，有助于血管成熟。总的来说，我们的数据表明，无血清培养基或低氧预处理增强了 hASC 分泌组在临床前伤口愈合模型中的血管再生作用。
