Santos Jerran, Milthorpe Bruce K, Herbert Benjamin R, Padula Matthew P
Advanced Tissue Regeneration & Drug Delivery Group, School of Life Sciences, University of Technology Sydney, NSW, Australia.
Proteomics Core Facility, School of Life Sciences, University of Technology Sydney, NSW, Australia.
Int J Stem Cells. 2017 Nov 30;10(2):193-217. doi: 10.15283/ijsc17036.
Adipose derived stem cells (ADSCs) are acquired from abdominal liposuction yielding a thousand fold more stem cells per millilitre than those from bone marrow. A large research void exists as to whether ADSCs are capable of transdermal differentiation toward neuronal phenotypes. Previous studies have investigated the use of chemical cocktails with varying inconclusive results.
Human ADSCs were treated with a chemical stimulant, beta-mercaptoethanol, to direct them toward a neuronal-like lineage within 24 hours. Quantitative proteomics using iTRAQ was then performed to ascertain protein abundance differences between ADSCs, beta-mercaptoethanol treated ADSCs and a glioblastoma cell line.
The soluble proteome of ADSCs differentiated for 12 hours and 24 hours was significantly different from basal ADSCs and control cells, expressing a number of remodeling, neuroprotective and neuroproliferative proteins. However toward the later time point presented stress and shock related proteins were observed to be up regulated with a large down regulation of structural proteins. Cytokine profiles support a large cellular remodeling shift as well indicating cellular distress.
The earlier time point indicates an initiation of differentiation. At the latter time point there is a vast loss of cell population during treatment. At 24 hours drastically decreased cytokine profiles and overexpression of stress proteins reveal that exposure to beta-mercaptoethanol beyond 24 hours may not be suitable for clinical application as our results indicate that the cells are in trauma whilst producing neuronal-like morphologies. The shorter treatment time is promising, indicating a reducing agent has fast acting potential to initiate neuronal differentiation of ADSCs.
脂肪来源干细胞（ADSCs）取自腹部抽脂术，每毫升产生的干细胞数量比骨髓来源的干细胞多一千倍。关于ADSCs是否能够经皮向神经元表型分化，存在很大的研究空白。先前的研究调查了使用化学混合物的情况，但结果不一。
用人ADSCs与化学刺激剂β-巯基乙醇处理，使其在24小时内定向分化为神经元样谱系。然后使用iTRAQ进行定量蛋白质组学分析，以确定ADSCs、经β-巯基乙醇处理的ADSCs和胶质母细胞瘤细胞系之间的蛋白质丰度差异。
分化12小时和24小时的ADSCs可溶性蛋白质组与基础ADSCs和对照细胞有显著差异，表达了许多重塑、神经保护和神经增殖蛋白。然而，在后期时间点，观察到与应激和休克相关的蛋白质上调，同时结构蛋白大量下调。细胞因子谱也支持细胞发生了大规模重塑转变，表明细胞处于应激状态。
早期时间点表明分化开始。在后期时间点，处理过程中细胞数量大量减少。在24小时时，细胞因子谱急剧下降和应激蛋白过度表达表明，超过24小时暴露于β-巯基乙醇可能不适合临床应用，因为我们的结果表明细胞在产生神经元样形态的同时处于创伤状态。较短的处理时间很有前景，表明一种还原剂具有快速启动ADSCs神经元分化的潜力。
