Release of adsorbed fibronectin from temperature-responsive culture surfaces requires cellular activity.
Release of adsorbed fibronectin from temperature-responsive culture surfaces requires cellular activity.
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DOI：
10.1016/s0142-9612(99)00239-2
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发表时间：
2000-05
期刊：
影响因子：
14
通讯作者：
M. Yamato;Chie Konno;A. Kushida;M. Hirose;M. Utsumi;A. Kikuchi;T. Okano
中科院分区：
文献类型：
--
作者：
M. Yamato;Chie Konno;A. Kushida;M. Hirose;M. Utsumi;A. Kikuchi;T. Okano
我们之前已经通过接枝聚(N-异丙基丙烯酰胺)开发了一种温度响应型细胞培养表面，该表面的疏水性随着温度的变化而改变。虽然该表面表现出与商用聚苯乙烯细胞培养表面相似的疏水性，并在37℃时促进细胞黏附和增殖，但接枝聚合物在32℃以下变得亲水性，并在没有胰酶的情况下释放散布的培养细胞。温度调节的细胞分离需要细胞代谢活动，需要ATP消耗、信号转导和细胞骨架重组。用纤维粘连蛋白(FN)预涂覆这些表面，可以改善粘附性较差的培养肝细胞的铺展，并降低培养温度，从FN吸附的移植表面释放培养细胞。抗FN抗体免疫染色显示，降温后仅有位于培养细胞下方的FN从培养表面去除。降温后，FN被吸附到没有直接细胞附着的表面区域，仍然是表面结合的。本文还讨论了活性细胞脱离的新概念。
We have previously developed a temperature-responsive cell culture surface by grafting poly(N-isopropylacrylamide) that changes its surface hydrophobicity in response to temperature. While this surface shows similar hydrophobicity to that of commercial polystyrene cell culture surfaces and facilitates cell adhesion and proliferation at 37°C, grafted polymer becomes hydrophilic below 32°C and releases spread cultured cells without trypsin. Temperature-regulated cell detachment requires cell metabolic activity requiring ATP consumption, signal transduction, and cytoskeleton reorganziation. Precoating these surfaces with fibronectin (FN) improves spreading of less adhesive cultured hepatocytes and reducing culture temperature releases cultured cells from FN-adsorbed grafted surfaces. Immunostaining with anti-FN antibody revealed that only FN located beneath cultured cells is removed from culture surfaces after reducing temperature. FN adsorbed to surface areas lacking direct cell attachment remained surface-bound after reducing temperature. A novel concept of active cell detachment is also discussed.