Steele J G, Johnson G, Underwood P A
Division of Biomolecular Engineering, Commonwealth Scientific and Industrial Research Organization, North Ryde, NSW, Australia.
J Biomed Mater Res. 1992 Jul;26(7):861-84. doi: 10.1002/jbm.820260704.
The suitability of polymeric biomaterials as surfaces for the attachment and growth of cells has often been investigated in tissue culture. In this study the contribution that adsorption of serum fibronectin (Fn) or vitronectin (Vn) make to the attachment and spreading of fibroblast cells during the first 90 min following seeding was determined for two modified tissue culture polystyrenes, as model biomaterial surfaces. The amount of serum Vn and Fn which adsorbed onto tissue culture grade polystyrene (TCP) from different serum concentrations over the range of 0.1-30% (v/v) were determined and compared to attachment of cells of the BHK-21 and HT1080 fibroblast lines. There was no simple correlation between the amount of Fn or the amount of Vn adsorbed and cell attachment and spreading. The requirement for Fn or Vn for attachment and spreading of BHK-21 or HT1080 cells onto modified polystyrene (either TCP or to Primaria) during the first 90 min of cell culture was directly tested by selective removal of Fn or Vn from the serum prior to addition to the culture medium. Attachment and spreading of BHK-21 or HT1080 cells onto TCP or Primaria surfaces were reduced in a concentration-dependent manner when the cells were seeded in medium containing 2% (v/v) or higher concentrations of Vn-depleted serum. BHK-21 cells or HT1080 cells seeded in medium containing Fn-depleted serum (which contained Vn) attached and spread onto TCP or Primaria. Both BHK-21 cells and HT1080 cells failed to attach to TCP or Primaria when seeded in medium containing serum depleted of both Vn and Fn. The requirement for serum Vn or Fn for fibroblast attachment to TCP was also tested using cells of a human dermal fibroblast strain. The attachment of the dermal fibroblasts to TCP during the first 90 min of culture was not decreased by depletion of Vn from the 15% (v/v) serum, but there was a reduction in the proportion of the attached cells which had spread. Selective depletion of serum Fn did not have any effect on either cell attachment or spreading. Our results show that for fibroblast cells, particularly with cell lines such as BHK-21 or HT1080 but also with cell strains, the first binding of cells onto tissue culture polystyrene when plated in medium containing serum is a result of adsorption onto the surface of serum Vn. The adsorption of serum Vn onto the surface overcomes the effect of serum components which tend to decrease cell attachment.
在组织培养中，人们经常研究聚合生物材料作为细胞附着和生长表面的适用性。在本研究中，对于两种经过改性的组织培养聚苯乙烯（作为模型生物材料表面），测定了血清纤连蛋白（Fn）或玻连蛋白（Vn）的吸附在接种后最初90分钟内对成纤维细胞附着和铺展的贡献。测定了在0.1 - 30%（v/v）范围内不同血清浓度下吸附到组织培养级聚苯乙烯（TCP）上的血清Vn和Fn的量，并将其与BHK - 21和HT1080成纤维细胞系的细胞附着情况进行比较。Fn或Vn的吸附量与细胞附着和铺展之间没有简单的相关性。通过在添加到培养基之前从血清中选择性去除Fn或Vn，直接测试了BHK - 21或HT1080细胞在细胞培养最初90分钟内附着和铺展到改性聚苯乙烯（TCP或Primaria）上对Fn或Vn的需求。当细胞接种在含有2%（v/v）或更高浓度Vn耗尽血清的培养基中时，BHK - 21或HT1080细胞在TCP或Primaria表面的附着和铺展以浓度依赖的方式减少。接种在含有Fn耗尽血清（含有Vn）的培养基中的BHK - 21细胞或HT1080细胞附着并铺展到TCP或Primaria上。当接种在含有同时耗尽Vn和Fn的血清的培养基中时，BHK - 21细胞和HT1080细胞均未能附着到TCP或Primaria上。还使用人皮肤成纤维细胞株的细胞测试了血清Vn或Fn对成纤维细胞附着到TCP的需求。在培养的最初90分钟内，从15%（v/v）血清中耗尽Vn并没有降低皮肤成纤维细胞对TCP的附着，但附着细胞铺展的比例有所降低。选择性耗尽血清Fn对细胞附着或铺展均没有任何影响。我们的结果表明，对于成纤维细胞，特别是对于诸如BHK - 21或HT1080这样的细胞系以及细胞株，当接种在含有血清的培养基中时，细胞首次结合到组织培养聚苯乙烯上是血清Vn吸附到表面的结果。血清Vn吸附到表面克服了倾向于降低细胞附着的血清成分的影响。
