Abstract:
This study aims to evaluate and compare cellular therapy with human Wharton\'s jelly (WJ) mesenchymal stem cells (MSCs) and neural precursors (NPs) in experimental autoimmune encephalomyelitis (EAE), a preclinical model of Multiple Sclerosis. MSCs were isolated from WJ by an explant technique, differentiated to NPs, and characterized by cytometry and immunocytochemistry analysis after ethical approval. Forty-eight rats were EAE-induced by myelin basic protein and Freund\'s complete adjuvant. Forty-eight hours later, the animals received intraperitoneal injections of 250 ng/dose of Bordetella pertussis toxin. Fourteen days later, the animals were divided into the following groups: a. non-induced, induced: b. Sham, c. WJ-MSCs, d. NPs, and e. WJ-MSCs plus NPs. 1 × 105. Moreover, the cells were placed in a 10 µL solution and injected via a stereotaxic intracerebral ventricular injection. After ten days, the histopathological analysis for H&E, Luxol, interleukins, and CD4/CD8 was carried out. Statistical analyses demonstrated a higher frequency of clinical manifestation in the Sham group (15.66%) than in the other groups; less demyelination was seen in the treated groups than the Sham group (WJ-MSCs, p = 0.016; NPs, p = 0.010; WJ-MSCs + NPs, p = 0.000), and a lower cellular death rate was seen in the treated groups compared with the Sham group. A CD4/CD8 ratio of <1 showed no association with microglial activation (p = 0.366), astrocytes (p = 0.247), and cell death (p = 0.577) in WJ-MSCs. WJ-MSCs and NPs were immunomodulatory and neuroprotective in cellular therapy, which would be translated as an adjunct in demyelinating diseases.
摘要:
本研究旨在评估和比较人沃顿果冻(WJ)间充质干细胞(MSCs)和神经前体(NPs)在实验性自身免疫性脑脊髓炎(EAE)中的细胞治疗,多发性硬化症的临床前模型。通过外植体技术从WJ中分离出MSCs,区分为NP,并在伦理批准后通过细胞计数和免疫细胞化学分析进行表征。48只大鼠由髓磷脂碱性蛋白和弗氏完全佐剂诱导EAE。48小时后,这些动物接受腹膜内注射250ng/剂百日咳博德特氏菌毒素.十四天后,将动物分为以下组:a.非诱导,诱导:b.Sham,c.WJ-MSC,d.NP,和e.WJ-MSC加NP。1×105此外,将细胞置于10µL溶液中,并通过立体定位脑室注射进行注射.十天后,H&E的组织病理学分析,Luxol,白细胞介素,进行CD4/CD8。统计分析表明,与其他组相比,Sham组的临床表现频率更高(15.66%);与Sham组相比,治疗组的脱髓鞘减少(WJ-MSCs,p=0.016;NPs,p=0.010;WJ-MSC+NP,p=0.000),与Sham组相比,治疗组的细胞死亡率较低。CD4/CD8比值<1显示与小胶质细胞活化无关(p=0.366),星形胶质细胞(p=0.247),和WJ-MSC中的细胞死亡(p=0.577)。WJ-MSCs和NPs在细胞治疗中具有免疫调节和神经保护作用,这将被翻译为脱髓鞘疾病的辅助手段。