Wu Szu-Hsien, Liao Yu-Ting, Hsueh Kuang-Kai, Huang Hui-Kuang, Chen Tung-Ming, Chiang En-Rung, Hsu Shan-Hui, Tseng Ting-Chen, Wang Jung-Pan
Division of Plastic and Reconstructive Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan.
Department of Surgery, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
Front Cell Dev Biol. 2021 Apr 30;9:658099. doi: 10.3389/fcell.2021.658099. eCollection 2021.
Hypoxic expansion has been demonstrated to enhance neuronal differentiation of bone-marrow derived mesenchymal stem cells (BMSCs). Whether adipose-derived mesenchymal stem cells (ADSCs) increase their neuronal differentiation potential following hypoxic expansion has been examined in the study. Real-time quantitative reverse transcription-polymerase chain reaction and immunofluorescence staining were employed to detect the expression of neuronal markers and compare the differentiation efficiency of hypoxic and normoxic ADSCs. A sciatic nerve injury animal model was used to analyze the gastrocnemius muscle weights as the outcomes of hypoxic and normoxic ADSC treatments, and sections of the regenerated nerve fibers taken from the conduits were analyzed by histological staining and immunohistochemical staining. Comparisons of the treatment effects of ADSCs and BMSCs following hypoxic expansion were also conducted and . Hypoxic expansion prior to the differentiation procedure promoted the expression of the neuronal markers in ADSC differentiated neuron-like cells. Moreover, the conduit connecting the sciatic nerve gap injected with hypoxic ADSCs showed the highest recovery rate of the gastrocnemius muscle weights in the animal model, suggesting a conceivable treatment for hypoxic ADSCs. The percentages of the regenerated myelinated fibers from the hypoxic ADSCs detected by toluidine blue staining and myelin basic protein (MBP) immunostaining were higher than those of the normoxic ones. On the other hand, hypoxic expansion increased the neuronal differentiation potential of ADSCs compared with that of the hypoxic BMSCs . The outcomes of animals treated with hypoxic ADSCs and hypoxic BMSCs showed similar results, confirming that hypoxic expansion enhances the neuronal differentiation potential of ADSCs and improves therapeutic potential.
缺氧扩增已被证明可增强骨髓间充质干细胞（BMSCs）的神经元分化。本研究探讨了脂肪来源的间充质干细胞（ADSCs）在缺氧扩增后其神经元分化潜能是否增加。采用实时定量逆转录-聚合酶链反应和免疫荧光染色检测神经元标志物的表达，并比较缺氧和常氧ADSCs的分化效率。利用坐骨神经损伤动物模型分析缺氧和常氧ADSCs治疗后的腓肠肌重量，并通过组织学染色和免疫组织化学染色分析取自导管的再生神经纤维切片。还对缺氧扩增后ADSCs和BMSCs的治疗效果进行了比较。分化程序前的缺氧扩增促进了ADSC分化的神经元样细胞中神经元标志物的表达。此外，在动物模型中，注射缺氧ADSCs的连接坐骨神经间隙的导管显示腓肠肌重量的恢复率最高，提示缺氧ADSCs可能是一种可行的治疗方法。通过甲苯胺蓝染色和髓鞘碱性蛋白（MBP）免疫染色检测到的缺氧ADSCs再生有髓纤维的百分比高于常氧ADSCs。另一方面，与缺氧BMSCs相比，缺氧扩增增加了ADSCs的神经元分化潜能。缺氧ADSCs和缺氧BMSCs治疗动物的结果显示相似的结果，证实缺氧扩增增强了ADSCs的神经元分化潜能并提高了治疗潜力。
