Groves B J, Riley P A
Dental School, University College and Middlesex School of Medicine, London, Great Britain.
Cytobios. 1987;52(208):49-62.
The design and construction of a miniaturised shearing apparatus is described. Cultures (24 h) of an established epithelial cell line (BEB) were exposed to flow conditions in the shearing chamber at 37 degrees C, and subsequently glass coverslip cultures were prepared for photography. The critical shear radius (CSR) was determined by densitometry from a negative film and the minimum distraction force (MDF) at the CSR calculated using predetermined viscosity values of the flow medium. The mean calculated MDF of BEB cells ranged from 1.04-1.36 Nm-2, and was independent of the culture inoculation density (9 to 37 X 10(4) cells cm-2) and the time (5-20 min) of exposure to shearing conditions. The MDF was increased to 3.3 Nm-2 by a 30 min exposure of cultures to concanavalin A (50 micrograms ml-1), and this effect was abolished by treating with mannose (0.05 M). The results demonstrated that the radial flow chamber principle is applicable to the measurement of cell to substratum adhesion of cultured mammalian cells.
本文描述了一种小型剪切装置的设计与构建。将已建立的上皮细胞系(BEB)培养24小时后,置于37摄氏度的剪切室中使其暴露于流动条件下,随后制备玻璃盖玻片培养物用于拍照。通过对负片进行光密度测定来确定临界剪切半径(CSR),并使用流动介质的预定粘度值计算CSR处的最小牵张力(MDF)。BEB细胞的平均计算MDF范围为1.04 - 1.36 Nm-2,且与培养接种密度(9至37×10(4)个细胞/cm2)以及暴露于剪切条件的时间(5 - 20分钟)无关。通过将培养物暴露于伴刀豆球蛋白A(50微克/毫升)30分钟,MDF增加至3.3 Nm-2,而用甘露糖(0.05 M)处理可消除此效应。结果表明,径向流动室原理适用于测量培养的哺乳动物细胞与基质的粘附力。