Ma Yee Hang Ethan, Putta Abhinay R, Chan Cyrus H H, Vidman Stephen R, Monje Paula, Plant Giles W
Department of Neuroscience and Chronic Brain Injury Program, The Ohio State University, Columbus, OH 43210, USA.
Department of Neuroscience, The Ohio State University, Columbus, OH 43210, USA.
Cells. 2025 Mar 20;14(6):461. doi: 10.3390/cells14060461.
Schwann cell (SC) transplantation shows promise in treating spinal cord injury as a pro-regenerative agent to allow host endogenous neurons to bridge over the lesion. However, SC transplants face significant oxidative stress facilitated by ROS in the lesion, leading to poor survival. deferoxamine mesylate (DFO) is a neuroprotective agent shown to reduce HO-induced cell death in serum-containing conditions. Here we show that DFO is not necessary to induce neuroprotection under serum-free conditions by cell survival quantification and phenotypic analysis via immunohistochemistry, Hif1α and collagen IV quantification via whole cell corrected total cell fluorescence, and cell death transcript changes via RT-qPCR. Our results indicate survival of SC regardless of DFO pretreatment in serum-free conditions and an increased survival facilitated by DFO in serum-containing conditions. Furthermore, our results showed strong nuclear expression of Hif1α in serum-free conditions regardless of DFO pre-treatment and a nuclear expression of Hif1α in DFO-treated SCs in serum conditions. Transcriptomic analysis reveals upregulation of autophagy transcripts in SCs grown in serum-free media relative to SCs in serum conditions, with and without DFO and HO. Thus, indicating a pro-repair and regenerative state of the SCs in serum-free conditions. Overall, results indicate the protectiveness of CDM in enhancing SC survival against ROS-induced cell death in vitro.
施万细胞（SC）移植作为一种促进再生的因子，有望用于治疗脊髓损伤，使宿主内源性神经元跨越损伤部位。然而，SC移植面临着损伤部位活性氧（ROS）引发的严重氧化应激，导致存活率低下。甲磺酸去铁胺（DFO）是一种神经保护剂，在含血清条件下可减少血红素加氧酶（HO）诱导的细胞死亡。在此，我们通过免疫组织化学进行细胞存活定量和表型分析、通过全细胞校正总细胞荧光进行Hif1α和IV型胶原定量以及通过逆转录定量聚合酶链反应（RT-qPCR）分析细胞死亡转录本变化，结果表明在无血清条件下诱导神经保护并不需要DFO。我们的结果表明，在无血清条件下，无论是否进行DFO预处理，SC均能存活，而在含血清条件下，DFO可提高其存活率。此外，我们的结果显示，在无血清条件下，无论是否进行DFO预处理，Hif1α均有强烈的核表达，而在含血清条件下经DFO处理的SC中，Hif1α也有核表达。转录组分析显示，相对于在含血清条件下培养的SC，无论有无DFO和HO，在无血清培养基中培养的SC中自噬转录本上调。因此，这表明在无血清条件下SC处于促进修复和再生的状态。总体而言，结果表明无血清培养基（CDM）在增强SC存活以抵抗ROS诱导的体外细胞死亡方面具有保护作用。
