Micropatterned immobilization of epidermal growth factor to regulate cell function
Micropatterned immobilization of epidermal growth factor to regulate cell function
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DOI：
10.1021/bc970190b
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发表时间：
1998-03-01
影响因子：
4.7
通讯作者：
Imanishi, Y
中科院分区：
文献类型：
--
作者：
Ito, Y;Chen, GP;Imanishi, Y
将光敏聚烯丙基胺与N-[4-（叠氮苯甲酰基）氧基]琥珀酰亚胺偶联得到光敏聚烯丙基胺，再与小鼠表皮生长因子（EGF）偶联得到光敏EGF。在光掩模的存在下，通过UV照射将EGF衍生物以规定的微图案图案固定在聚苯乙烯板上。EGF在聚苯乙烯板上的图案化固定通过用抗EGF抗体的免疫染色来确认。将过表达EGF受体的中国仓鼠卵巢（CHO）细胞在微图案化板上培养。信号蛋白，包括EGF受体的磷酸化酪氨酸残基，仅在EGF固定区域中粘附的细胞中检测到，并且仅在EGF固定区域中观察到细胞生长。当EGF固定面积小于细胞面积时，抗磷酸酪氨酸抗体对生长在EGF固定区域的细胞部分染色。活化蛋白的部分染色表明EGF的固定抑制了活化的EGF-EGF受体复合物的自由侧向扩散和内化。增强的细胞生长是由于通过固定的EGF实现的阿曲他克林刺激。
Photoreactive epidermal growth factor (EGF) was synthesized by conjugating mouse EGF with photoreactive polyallylamine, which was synthesized by the coupling reaction of polyallylamine with N-[4-(azidobenzoyl)oxy]succinimide. The EGF derivative was pattern-immobilized onto a polystyrene plate by UV irradiation in the presence of a photomask in a prescribed micropattern. The patterned immobilization of EGF on the polystyrene plate was confirmed by immunostaining with anti-EGF antibody. Chinese hamster ovary (CHO) cells overexpressing EGF receptors were cultured on the micropatterned plate. The phosphorylated tyrosine residues of signal proteins, including EGF receptors, were detected only in the cells adhered in the EGF-immobilized area, and cell growth was observed only in the EGF-immobilized area. The cells growing in the EGF-immobilized area were partially stained by anti-phosphotyrosine antibody, when the area of EGF immobilization was smaller than the cell. The partial staining of activated proteins indicates that immobilization of EGF inhibited the free lateral diffusion and internalization of the activated EGF-EGF receptor complex. The enhanced cell growth is due to juxtacrine stimulation realized by immobilized EGF.