Stem cells from human exfoliated deciduous teeth (SHEDs) are highly proliferative, clonogenic, and multipotent stem cells with a neural crest cell origin. This property could be a desirable option for potential therapeutic applications. In this study, we focus on the effects of Rho kinase inhibitors Y‐27632 and Noggin on the proliferation of SHEDs and their differentiation into neuron‐like cells. SHEDs were extracted from 10 samples of deciduous teeth obtained from healthy children aged from 5 to 10. The passaged SHEDs were transfected with Noggin, Y‐27632, or their combination. By means of MTT and colony formation assays, the effects of Y‐27632 and Noggin on cell viability and colony formation were detected. Cellular morphology and neurosphere formation were observed under a microscope. Y‐27632 transfection in SHEDs showed enhanced cell viability, colony formation, and neurosphere formation indicating that Y‐27632 could promote cell proliferation of SHEDs. Furthermore, we observed that the SHEDs treated with Noggin in combination with Y‐27632 displayed typical neuron‐like cell morphology and reticular processes. Noggin or Y‐27632 alone or in combination induced obviously increased NSE, Nestin, and GFAP levels, which were highest in SHEDs treated with the combination of Noggin and Y‐27632. These findings suggest that Y‐27632 promotes the proliferation of SHEDs, and Y‐27632 and Noggin in combination have a synergistic effect on promoting differentiation of SHEDs into neuron‐like cells.

中文翻译：

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Rho激酶抑制剂Y-27632和Noggin过表达对人脱落乳牙来源干细胞增殖和神经元样细胞分化的协同作用

来自人类脱落乳牙 (SHED) 的干细胞是具有神经嵴细胞起源的高度增殖性、克隆性和多能干细胞。这种特性可能是潜在治疗应用的理想选择。在这项研究中，我们关注 Rho 激酶抑制剂 Y-27632 和 Noggin 对 SHED 增殖及其分化为神经元样细胞的影响。从 5 至 10 岁健康儿童的 10 个乳牙样本中提取 SHED。用 Noggin、Y-27632 或它们的组合转染传代的 SHED。通过 MTT 和集落形成试验，检测了 Y-27632 和 Noggin 对细胞活力和集落形成的影响。在显微镜下观察细胞形态和神经球形成。SHED 中的 Y-27632 转染显示细胞活力增强，集落形成和神经球形成表明 Y-27632 可以促进 SHED 的细胞增殖。此外，我们观察到用 Noggin 与 Y-27632 组合处理的 SHED 显示出典型的神经元样细胞形态和网状突起。单独或联合使用 Noggin 或 Y-27632 可显着增加 NSE、Nestin 和 GFAP 水平，在用 Noggin 和 Y-27632 组合处理的 SHED 中最高。这些发现表明Y-27632促进SHEDs的增殖，Y-27632和Noggin联合使用对促进SHEDs向神经元样细胞分化具有协同作用。我们观察到用 Noggin 与 Y-27632 组合处理的 SHED 显示出典型的神经元样细胞形态和网状突起。单独或联合使用 Noggin 或 Y-27632 可显着增加 NSE、Nestin 和 GFAP 水平，在用 Noggin 和 Y-27632 组合处理的 SHED 中最高。这些发现表明Y-27632促进SHEDs的增殖，Y-27632和Noggin联合使用对促进SHEDs向神经元样细胞分化具有协同作用。我们观察到用 Noggin 与 Y-27632 组合处理的 SHED 显示出典型的神经元样细胞形态和网状突起。Noggin 或 Y-27632 单独或联合诱导显着增加 NSE、Nestin 和 GFAP 水平，这在用 Noggin 和 Y-27632 联合处理的 SHED 中最高。这些发现表明Y-27632促进SHEDs的增殖，Y-27632和Noggin联合使用对促进SHEDs向神经元样细胞分化具有协同作用。