Chin Sze-Piaw, Abd Rahim Erlena Nor Asmira, Nor Arfuzir Natasha Najwa
Cytopeutics Sdn Bhd, Bio-X Centre, Persiaran Cyberpoint Selatan, Cyber 8, 63000, Cyberjaya, Selangor, Malaysia.
CMH Specialist Hospital, Jalan Tun Dr. Ismail, 70200, Seremban, Negeri Sembilan, Malaysia.
In Vitro Cell Dev Biol Anim. 2025 May 13. doi: 10.1007/s11626-025-01037-y.
Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are a potential off-the-shelf product for acute ischemic stroke. This study explored the underlying mechanism of Cytopeutics® hUC-MSCs (Neuroncell-EX) as well as its feasibility and efficacy at two different doses: 2 × 10 cells per rat and 4 × 10 cells/rat in middle cerebral artery occlusion (MCAO) ischemic stroke model for 28 d. Modified neurological severity score (mNSS) and rotarod tests were evaluated at days 1, 4, 7, and 14. Transforming growth factor-beta 1 (TGF-β1), interleukin-1 receptor antagonist (IL-1Ra), and vascular endothelial growth factor (VEGF) were evaluated by enzyme-linked immunosorbent assay (ELISA) at days 4 and 28. Immunohistochemistry expression of aquaporin-4 (AQP4) and neuronal protein marker (NeuN) were performed at days 4 and 28, respectively. Both doses of Neuroncell-EX showed significant lower mNSS scores at days 7 and 14 compared to stroke control. Both Neuroncell-EX groups showed significant longer latency time at day 7, with only 4 × 10⁶ cells/rat group having significant longer time at day 14 than stroke control. At both time points, the 2 × 10⁶ cells/rat group had significantly higher TGF-β1 and IL-1Ra levels, with significantly increased TGF-β1 only observed in 4 × 10⁶ cells/rat group at day 4 compared to stroke control. The VEGF levels were significantly lower at day 4 but then significantly increased at day 28 in both Neuroncell-EX groups than stroke control. AQP4 expression was significantly higher in stroke control compared to healthy control at day 4. Both doses of Neuroncell-EX showed significantly higher NeuN expression compared to stroke control at day 28. There is a weak correlation between TGF-β1 with VEGF and inversely with AQP4. These results suggest that Neuroncell-EX is feasible and effective in promoting functional recovery and neuroprotection in ischemic rats, potentially through immunomodulation, angiogenesis, and neurogenesis mechanisms.
人脐带间充质干细胞（hUC-MSCs）是急性缺血性中风潜在的现成可用产品。本研究探讨了Cytopeutics® hUC-MSCs（Neuroncell-EX）的潜在机制及其在两种不同剂量下的可行性和疗效：在大脑中动脉闭塞（MCAO）缺血性中风模型中，每只大鼠分别给予2×10⁶个细胞和4×10⁶个细胞，持续28天。在第1、4、7和14天评估改良神经功能缺损评分（mNSS）和转棒试验。在第4天和第28天通过酶联免疫吸附测定（ELISA）评估转化生长因子-β1（TGF-β1）、白细胞介素-1受体拮抗剂（IL-1Ra）和血管内皮生长因子（VEGF）。分别在第4天和第28天进行水通道蛋白-4（AQP4）和神经元蛋白标志物（NeuN）的免疫组织化学表达检测。与中风对照组相比，两种剂量的Neuroncell-EX在第7天和第14天的mNSS评分均显著降低。两个Neuroncell-EX组在第7天的潜伏期均显著延长，只有每只大鼠给予4×10⁶个细胞的组在第14天的时间比中风对照组显著延长。在两个时间点，每只大鼠给予2×10⁶个细胞的组的TGF-β1和IL-1Ra水平均显著更高，与中风对照组相比，仅在每只大鼠给予4×10⁶个细胞的组在第4天观察到TGF-β1显著增加。与中风对照组相比，两个Neuroncell-EX组在第4天的VEGF水平显著降低，但在第28天显著升高。与健康对照组相比，中风对照组在第4天的AQP4表达显著更高。与中风对照组相比，两种剂量的Neuroncell-EX在第28天的NeuN表达均显著更高。TGF-β1与VEGF之间存在弱相关性，与AQP4呈负相关。这些结果表明，Neuroncell-EX在促进缺血大鼠的功能恢复和神经保护方面是可行且有效的，可能是通过免疫调节、血管生成和神经发生机制实现的。
