Differentiation of Mesenchymal Stem Cells Derived from Amniotic Membrane to Neuronal Cells and the expression of PAX2 and NURR1 Genes

Nooshin Barikrow, Javaneh Khosravi
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Abstract

: Mesenchymal stem cells (MSCs) have different sources, including bone marrow, adipose tissue, umbilical cord, amniotic fluid, and amniotic membrane. They have immunomodulatory properties. These cells can be used for the treatment of many neurological diseases such as Parkinson’s and Alzheimer’s disease. In this study, MSCs were isolated from the amniotic membrane, and their surface markers were investigated using flow cytometry. MSCs were differentiated to osteoblasts, adipocytes, and finally, to the nerve cells under the influence of epidermal, basic fibroblast natural growth factors, and two other media. The first medium included indomethacin, butyric acid, and ascorbic acid. The second one included retinoic acid and ascorbic acid. The expression of paired box gene 2 (PAX2) and nuclear receptor related-1 (NURR1) genes were investigated using real-time polymerase chain reaction and showed higher levels than that of controls. The presence of the expression of β-tubulin III and microtubule-associated protein 2 (MAPII) was studied by immunocytochemistry. The result suggested that the second medium included retinoic acid was better than the first medium. Protecting neurons are important in neurological diseases, and the expression of mRNAs such as PAX2, NURR1, β-tubulin III, and MAPII plays an effective role in neural differentiation.
羊膜间充质干细胞向神经细胞的分化及PAX2和NURR1基因的表达
间充质干细胞有不同的来源,包括骨髓、脂肪组织、脐带、羊水和羊膜。它们具有免疫调节特性。这些细胞可用于治疗许多神经系统疾病,如帕金森病和阿尔茨海默病。本研究从羊膜中分离MSCs,用流式细胞术检测其表面标记物。MSCs在表皮、碱性成纤维细胞自然生长因子和其他两种培养基的作用下分化为成骨细胞、脂肪细胞,最后分化为神经细胞。第一种培养基包括吲哚美辛、丁酸和抗坏血酸。第二种包括维甲酸和抗坏血酸。实时聚合酶链反应检测配对盒基因2 (PAX2)和核受体相关-1 (NURR1)基因的表达水平均高于对照组。免疫细胞化学检测β-微管蛋白III和微管相关蛋白2 (MAPII)的表达。结果表明,含维甲酸的第二种培养基效果优于第一种培养基。保护神经元在神经系统疾病中具有重要意义,PAX2、NURR1、β-微管蛋白III、MAPII等mrna的表达在神经分化中起着有效的作用。
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