Mesenchymal stem cells (MSCs), and within them adipose tissue derived stem cells (ASCs), have been shown to have therapeutic effects on central nervous system (CNS) cell populations. Such effects have been mostly attributed to soluble factors, as well as vesicles, present in their secretome. Yet, little is known about the impact that MSC passaging might have in the secretion therapeutic profile. Our aim was to show how human ASCs (hASCs) passage number influences the effect of their secretome in neuronal survival, differentiation and axonal growth. For this purpose, post-natal rat hippocampal primary cultures, human neural progenitor cell (hNPCs) cultures and dorsal root ganglia (DRGs) explants were incubated with secretome, collected as conditioned media (CM), obtained from hASCs in P3, P6, P9 and P12. Results showed no differences when comparing percentages of MAP-2 positive cells (a mature neuronal marker) in neuronal cultures or hNPCs, after incubation with hASCs secretome from different passages. The same was observed regarding DRG neurite outgrowth. In order to characterize the secretomes obtained from different passages, a proteomic analysis was performed, revealing that its composition did not vary significantly with passage number P3 to P12. Results allowed us to identify several key proteins, such as pigment epithelium derived factor (PEDF), DJ-1, interleucin-6 (IL-6) and galectin, all of which have already proven to play neuroprotective and neurodifferentiating roles. Proteins that promote neurite outgrowth were also found present, such as semaphorin 7A and glypican-1. We conclude that cellular passaging does not influence significantly hASCs's secretome properties especially their ability to support post-natal neuronal survival, induce neurodifferentiation and promote axonal growth.

间充质干细胞（MSC）以及其中的脂肪组织衍生干细胞（ASC）已显示对中枢神经系统（CNS）细胞群具有治疗作用。这种作用主要归因于其分泌组中存在的可溶性因子以及囊泡。然而，关于MSC传代可能对分泌治疗概况的影响知之甚少。我们的目的是显示人类ASC（hASC）的传代次数如何影响其分泌组对神经元存活，分化和轴突生长的影响。为此，将产后大鼠海马原代培养物，人神经祖细胞（hNPCs）培养物和背根神经节（DRGs）外植体与分泌液一起孵育，并作为条件培养基（CM）收集，并从hASC中以P3，P6，P9的形式获得和P12。与来自不同传代的hASC分泌基因组孵育后，比较神经元培养物或hNPC中MAP-2阳性细胞（成熟的神经元标记）的百分比时，结果没有差异。对于DRG神经突向外生长，观察到相同的结果。为了表征从不同传代获得的分泌组，进行了蛋白质组学分析，揭示了其组成随传代数P3至P12没有明显变化。结果使我们能够鉴定出几种关键蛋白，例如色素上皮衍生因子（PEDF），DJ-1，白细胞介素6（IL-6）和半乳凝素，所有这些蛋白均已被证明具有神经保护作用和神经分化作用。还发现了促进神经突生长的蛋白质，例如信号蛋白7A和glypican-1。