Sina Trauma and Surgery Research Center, Sina Hospital, Tehran University of Medical Sciences, Hasan-Abad Square, Imam Khomeini Ave., Tehran, 11365-3876, Iran.
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Number 88, Italy Street, Between Ghods Street and Vesal Shirazi Street, Tehran, 14177-55469, Iran.
Cell Biol Int. 2020 May;44(5):1168-1183. doi: 10.1002/cbin.11315. Epub 2020 Feb 18.
Microtubule-stabilizing agents (MSAs), until now, have primarily been considered for their anti-proliferative effects in the setting of cancer. However, recent studies have revealed that one particular MSA, epothilone B (EpoB), can promote axonal regeneration after traumatic spinal cord injuries (SCI) even in the presence of inhibitor molecules such as neurite outgrowth inhibitor-A (Nogo-A). On the basis of the importance of having an efficient motor neuron (MN) differentiation protocol for stem cell therapy and the attention of MSAs for SCI treatment, our study investigated the effect of EpoB on human endometrial stem cells (hEnSCs) differentiation into MN-like cells. hEnSCs were isolated and characterized by flow cytometry. The hEnSC cell viability was evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To mimic the in vivo inhibitory environment, hEnSCs were also differentiated in the presence of Nogo-A. After 15 days of differentiation, the expressions of MN-markers were evaluated by real-time reverse-transcriptase polymerase chain reaction and immunofluorescence. According to the MTT assay results, three doses (1, 5, and 10 nM) of EpoB were selected to evaluate their effect on MN-differentiation. All selected doses can increase the efficacy of hEnSCs differentiation into MN-like cells. In particular, the 10 nM EpoB dosage was shown to increase the axon elongation, cell alignment, and upregulation of these MN-markers compared with other doses. EpoB can improve MN differentiation from hEnSC and potentially provide a unique route for neuronal replacement in the setting of SCI.
微管稳定剂（MSAs）迄今为止，主要因其在癌症中的抗增殖作用而被考虑。然而，最近的研究表明，一种特殊的 MSA，表鬼臼毒素 B（EpoB），即使在神经生长抑制因子-A（Nogo-A）等抑制分子存在的情况下，也能促进创伤性脊髓损伤（SCI）后的轴突再生。基于对干细胞治疗中有效运动神经元（MN）分化方案的重要性以及 MSAs 对 SCI 治疗的关注，我们的研究调查了 EpoB 对人子宫内膜干细胞（hEnSCs）分化为 MN 样细胞的影响。通过流式细胞术分离和鉴定 hEnSCs。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法评估 hEnSC 细胞活力。为了模拟体内抑制环境，还在存在 Nogo-A 的情况下对 hEnSCs 进行分化。分化 15 天后，通过实时逆转录聚合酶链反应和免疫荧光评估 MN 标志物的表达。根据 MTT 测定结果，选择三个剂量（1、5 和 10 nM）EpoB 来评估其对 MN 分化的影响。所有选定的剂量都可以提高 hEnSCs 分化为 MN 样细胞的效果。特别是，与其他剂量相比，10 nM 的 EpoB 剂量显示出增加轴突伸长、细胞排列和这些 MN 标志物的上调。EpoB 可以改善 hEnSC 中的 MN 分化，并为 SCI 中神经元替代提供独特途径。
