Developmental Biology Laboratory, Department of Animal Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
Department of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
J Cell Biochem. 2022 Jun;123(6):1120-1129. doi: 10.1002/jcb.30269. Epub 2022 May 9.
Permanent degeneration and loss of dopaminergic (DA) neurons in substantia nigra is the main cause of Parkinson's disease. Considering the therapeutic application of stem cells in neurodegeneration, we sought to examine the neurogenic differentiation potential of the newly introduced neural crest originated mesenchymal stem cells (MSCs), namely, trabecular meshwork-derived mesenchymal stem cells (TM-MSCs) compared to two other sources of MSCs, adipose tissue-derived stem cells (ADSCs) and bone marrow-derived mesenchymal stem cells (BM-MSCs). The three types of cells were therefore cultured in the presence and absence of a neural induction medium followed by the analysis of their differentiation potentials. Our results showed that TM-MSCs exhibited enhanced neural morphologies as well as higher expressions of MAP2 as the general neuron marker and Nurr-1 as an early DA marker compared to the adipose tissue-derived mesenchymal stem cells (AD-MSCs) and bone marrow-derived stem cells (BMSCs). Also, analysis of Nurr-1 immunostaining showed more intense Nurr-1 stained nuclei in the neurally induced TM-MSCs compared to those in the AD-MSCs, BMSCs, and noninduced control TM-MSCs. To examine if Wnt/beta-catenin pathway drives TM-MSCs towards a DA fate, we treated them with the Wnt agonist (CHIR, 3 μM) and the Wnt antagonist (IWP-2, 3 μM). Our results showed that the expressions of Nurr-1 and MAP2, as well as the Wnt/beta-catenin target genes, c-Myc and Cyclin D1, were significantly increased in the CHIR-treated TM-MSCs, but significantly reduced in those treated with IWP-2. Altogether, we declare first a higher neural potency of TM-MSCs compared to the more commonly used MSCs, BMSCs and ADSCs, and second that Wnt/beta-catenin activation directs the neurally induced TM-MSCs towards a DA fate.
中脑黑质多巴胺能（DA）神经元的永久性退化和丧失是帕金森病的主要原因。考虑到干细胞在神经退行性疾病中的治疗应用，我们试图研究新引入的神经嵴起源的间充质干细胞（MSCs），即脉络膜衍生的间充质干细胞（TM-MSCs）与其他两种来源的 MSCs，即脂肪组织衍生的干细胞（ADSCs）和骨髓衍生的间充质干细胞（BM-MSCs）的神经发生分化潜能。因此，在存在和不存在神经诱导培养基的情况下培养这三种类型的细胞，然后分析它们的分化潜能。我们的结果表明，与脂肪组织衍生的间充质干细胞（AD-MSCs）和骨髓衍生的间充质干细胞（BMSCs）相比，TM-MSCs 表现出增强的神经形态以及更高的 MAP2 表达作为一般神经元标志物和 Nurr-1 作为早期 DA 标志物。此外，Nurr-1 免疫染色分析显示，与 AD-MSCs、BMSCs 和未诱导的对照 TM-MSCs 相比，神经诱导的 TM-MSCs 中 Nurr-1 染色核更强烈。为了研究 Wnt/β-catenin 途径是否使 TM-MSCs 向 DA 命运分化，我们用 Wnt 激动剂（CHIR，3 μM）和 Wnt 拮抗剂（IWP-2，3 μM）处理它们。我们的结果表明，Nurr-1 和 MAP2 的表达以及 Wnt/β-catenin 靶基因 c-Myc 和 Cyclin D1 的表达在 CHIR 处理的 TM-MSCs 中显著增加，而在 IWP-2 处理的 TM-MSCs 中显著降低。总之，我们首次声明 TM-MSCs 比更常用的 MSCs、BMSCs 和 ADSCs 具有更高的神经潜能，其次，Wnt/β-catenin 激活使神经诱导的 TM-MSCs 向 DA 命运分化。
