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发育阶段依赖性神经干细胞对不同生物功能表面氯化甲基汞的敏感性。

Developmental stage dependent neural stem cells sensitivity to methylmercury chloride on different biofunctional surfaces.

机构信息

Mossakowski Medical Research Centre, Polish Academy of Sciences, NeuroRepair Department, Warsaw, Poland.

出版信息

Toxicol In Vitro. 2014 Feb;28(1):76-87. doi: 10.1016/j.tiv.2013.06.023. Epub 2013 Jul 17.

Abstract

Sensitivity of neural stem cells viability, proliferation and differentiation upon exposure to methylmercury chloride (MeHgCl) was investigated on different types of biofunctional surfaces. Patterns of biodomains created by microprinting/microspotting of poly-l-lysine or extracellular matrix proteins (fibronectin and vitronectin) allowed for non-specific electrostatic or specific, receptor mediated interactions, respectively, between stem cells and the surface. The neural stem cell line HUCB-NSC has been previously shown to be susceptible to MeHgCl in developmentally dependent manner. Here we demonstrated that developmental sensitivity of HUCB-NSC to MeHgCl depends upon the type of adhesive biomolecules and the geometry of biodomains. Proliferation of HUCB-NSC was diminished in time and MeHgCl concentration dependent manner. In addition, the response to MeHgCl was found to be cell-type dependent. Undifferentiated cells were the most sensitive independently of the type of bioactive domain. Significant decrease of GFAP+ cells was detected among cells growing on poly-l-lysine, while on fibronectin and vitronectin, this effect was observed only in the highest (1μM) concentration of MeHgCl. β-Tubulin III expressing cells were most sensitive on fibronectin domains. In addition, limited bioactive domains to μm in size, as compared to non-patterned larger area of the same adhesive substrate, exerted protective role. Thus, the surface area and type of cell/biofunctional surface interaction exerted significant influence on developmental stage and cell-type specific response of HUCB-NSC to MeHgCl.

摘要

研究了神经干细胞在暴露于氯化甲基汞(MeHgCl)时对不同类型生物功能表面的活力、增殖和分化的敏感性。通过微印刷/微点聚赖氨酸或细胞外基质蛋白(纤连蛋白和 vitronectin)图案的生物域的创建,允许干细胞和表面之间分别进行非特异性静电或特异性、受体介导的相互作用。先前已经表明神经干细胞系 HUCB-NSC 以发育依赖的方式易受 MeHgCl 影响。在这里,我们证明了 HUCB-NSC 对 MeHgCl 的发育敏感性取决于粘附生物分子的类型和生物域的几何形状。HUCB-NSC 的增殖以时间和 MeHgCl 浓度依赖的方式减少。此外,发现对 MeHgCl 的反应取决于细胞类型。无论生物活性域的类型如何,未分化细胞都是最敏感的。在聚赖氨酸上生长的细胞中检测到 GFAP+细胞的显著减少,而在纤连蛋白和 vitronectin 上,仅在 MeHgCl 的最高(1μM)浓度下观察到这种效应。β-微管蛋白 III 表达细胞在纤连蛋白域上最敏感。此外,与相同粘附底物的非图案较大区域相比,尺寸限制在 μm 内的有限生物活性域发挥了保护作用。因此,表面面积和细胞/生物功能表面相互作用的类型对 HUCB-NSC 对 MeHgCl 的发育阶段和细胞类型特异性反应产生了重大影响。

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