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应激聚苯乙烯会增加贴壁细胞对流体剪切力的膜敏感性:技术说明。

Stressed polystyrene causes increased membrane sensitivity of adherent cells to fluid shear force: technical note.

作者信息

Lau J Y, Tchao R

机构信息

Division of Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of the Sciences in Philadelphia, Philadelphia, PA 19104, USA.

出版信息

Eur Cell Mater. 2007 Sep 10;14:40-3; discussion 43-4. doi: 10.22203/ecm.v014a04.

DOI:10.22203/ecm.v014a04
PMID:17828704
Abstract

Adherent cells transduce signals from the extracellular matrix, which result in changes to various cell functions, including cell spreading and morphology. However, changes to mechanical properties of cell membranes due to adherence to a substratum have not been studied. Adherent Nara Bladder Tumour (NBT) II cells on polystyrene (PS) discs made from pure, atactic polystyrene react differently between the peripheral 1mm zone and centre of the disc. After application of a fluid shear force, cells on the peripheral zone resulted in 91.1-/+0.8% cell death due to instantaneous rupture of apical cell membrane, as determined by the Live/Dead cell assay, whereas cells on the disc's centre and surrounding glass surface showed 7.1-/+5.7% and 4.3-/+1.7% cell death, respectively. Under cross-polarized light, the edge of the PS disc showed a low degree of birefringence whereas the centre of the disc did not. We also detached the PS disc and applied various weights (0.0 g to 40 g) to the disc at 100 degrees C for 2 hours and then cooled rapidly at 4 degrees C. We found that birefringence developed with stress to PS. NBT II cells grown on stressed PS showed an average of 55.8-/+14.1% cell death after a fluid shear force while cells on the glass surfaces resulted in only 5.0-/+2.7% cell death. Interestingly, increased birefringence is associated with increased lipophilicity on stressed PS, as determined by Nile Red staining. We propose that NBT II cell interaction with certain molecular characteristics of stressed PS results in altered cell membrane sensitivity to mechanical forces.

摘要

贴壁细胞可转导来自细胞外基质的信号,这些信号会导致多种细胞功能发生变化,包括细胞铺展和形态改变。然而,由于细胞贴附于基质而导致的细胞膜力学性质变化尚未得到研究。在由纯无规聚苯乙烯制成的聚苯乙烯(PS)圆盘上培养的奈良膀胱肿瘤(NBT)II贴壁细胞,在圆盘周边1mm区域和中心之间表现出不同的反应。施加流体剪切力后,通过活/死细胞检测法确定,周边区域的细胞由于顶端细胞膜瞬间破裂而导致91.1±0.8%的细胞死亡,而圆盘中心和周围玻璃表面的细胞死亡率分别为7.1±5.7%和4.3±1.7%。在交叉偏振光下,PS圆盘的边缘显示出低双折射,而圆盘中心则没有。我们还将PS圆盘分离,并在100℃下对圆盘施加各种重量(0.0g至40g),持续2小时,然后在4℃下快速冷却。我们发现PS在受力时会产生双折射。在受力的PS上生长的NBT II细胞在受到流体剪切力后平均有55.8±14.1%的细胞死亡,而在玻璃表面的细胞死亡率仅为5.0±2.7%。有趣的是,通过尼罗红染色确定,双折射增加与受力PS上的亲脂性增加有关。我们认为NBT II细胞与受力PS的某些分子特征相互作用会导致细胞膜对机械力的敏感性改变。

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