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两种诱导方案对人脂肪来源干细胞神经分化稳定性的影响。

Stability of neural differentiation in human adipose derived stem cells by two induction protocols.

机构信息

Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Tissue Cell. 2012 Apr;44(2):87-94. doi: 10.1016/j.tice.2011.11.006. Epub 2011 Dec 16.

Abstract

There are some evidences for suggesting that adipose derived stem cells (ADSCs) can be differentiated to the fate of neural cell type. ADSCs can be expanded rapidly in vitro and can be obtained by a less invasive method. In this study, we attempted to compare the stability of neural differentiation in human ADSCs by using two induction protocols. Isolated ADSCs were induced into neural-like cells using diverse effects of two specific procedures. For protocol 1, ADSCs were induced by chemical induction. In protocol 2, ADSCs were treated for sphere formation. Then, the singled cells were cultured in neurobasal media supplemented with special components. Differentiated ADSCs were evaluated for Nestin, MAP2 and GFAP expression by immunocytochemistry and semi quantitative RT-PCR techniques. Moreover, MTT assay was employed to detect cell viability and proliferation. Immunocytochemical analysis of both protocols demonstrated that ADSCs had large expression of the neural-specific markers. In RT-PCR, protocol 1 showed the highest percentage of MAP2 expression, but with time passing, the neural like state was reversible. Protocol 2 found with express of Nestin at week 1, however MAP2 and GFAP expression increased after 3 weeks. The neural-like cells produced by protocol 1 led to the further cell death. Comparative analysis showed that neural-like cell differentiation of ADSCs in chemical induction protocol was rapid but transitory, while it was approximately steady in neurosphere formation protocol.

摘要

有一些证据表明脂肪来源的干细胞(ADSCs)可以分化为神经细胞类型。ADSCs 可以在体外快速扩增,并可以通过一种微创的方法获得。在这项研究中,我们试图通过使用两种诱导方案来比较人 ADSCs 神经分化的稳定性。通过两种特定程序的不同作用将分离的 ADSCs 诱导为类神经细胞。方案 1 中,ADSCs 通过化学诱导进行诱导。在方案 2 中,ADSCs 经过球体形成处理。然后,将单细胞培养在含有特殊成分的神经基础培养基中。通过免疫细胞化学和半定量 RT-PCR 技术检测巢蛋白、MAP2 和 GFAP 的表达来评估分化的 ADSCs。此外,还采用 MTT 测定法检测细胞活力和增殖。两种方案的免疫细胞化学分析均表明 ADSCs 具有大量的神经特异性标志物表达。在 RT-PCR 中,方案 1 显示 MAP2 表达的百分比最高,但随着时间的推移,类神经状态是可逆的。方案 2 在第 1 周发现巢蛋白表达,但在第 3 周后 MAP2 和 GFAP 表达增加。方案 1 产生的类神经细胞导致进一步的细胞死亡。比较分析表明,化学诱导方案中 ADSCs 的类神经细胞分化快速但短暂,而神经球形成方案中的分化则相对稳定。

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