ROLE OF SERUM VITRONECTIN AND FIBRONECTIN IN ADHESION OF FIBROBLASTS FOLLOWING SEEDING ONTO TISSUE-CULTURE POLYSTYRENE
ROLE OF SERUM VITRONECTIN AND FIBRONECTIN IN ADHESION OF FIBROBLASTS FOLLOWING SEEDING ONTO TISSUE-CULTURE POLYSTYRENE
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DOI：
10.1002/jbm.820260704
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发表时间：
1992-07-01
期刊：
影响因子：
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通讯作者：
UNDERWOOD, PA
中科院分区：
文献类型：
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作者：
STEELE, JG;JOHNSON, G;UNDERWOOD, PA
高分子生物材料作为细胞附着和生长表面的适宜性在组织培养中经常被研究。在本研究中，测定了两种改性组织培养聚苯乙烯作为模型生物材料表面，血清纤维连接蛋白（Fn）或玻璃体连接蛋白（Vn）在播种后90分钟内对成纤维细胞附着和扩散的贡献。测定了在0.1 ~ 30% （v/v）范围内不同浓度的血清中Vn和Fn吸附在组织培养级聚苯乙烯（TCP）上的量，并与BHK-21和HT1080成纤维细胞系细胞的附着量进行了比较。Fn和Vn的吸附量与细胞附着和扩散之间没有简单的相关性。在细胞培养的前90分钟，通过在加入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细胞附着和扩散到改性聚苯乙烯（TCP或Primaria）上所需的Fn或Vn。当将BHK-21或HT1080细胞接种于含有2% （v/v）或更高浓度vn -贫血清的培养基中时，细胞在TCP或原代表面上的附着和扩散呈浓度依赖性减少。将BHK-21细胞或HT1080细胞接种到含有去氟血清（含Vn）的培养基中，附着并扩散到TCP或原代细胞上。BHK-21细胞和HT1080细胞在血清中同时缺失Vn和Fn时，均不能附着在TCP或原代细胞上。用人真皮成纤维细胞株的细胞也测试了成纤维细胞附着在TCP上所需的血清Vn或Fn。在培养的前90分钟内，皮肤成纤维细胞与TCP的附着并没有因为15% （v/v）血清中Vn的消耗而减少，但附着细胞扩散的比例有所减少。选择性去除血清Fn对细胞附着和扩散均无影响。我们的研究结果表明，对于成纤维细胞，特别是像BHK-21或HT1080这样的细胞系，以及细胞株，当细胞被镀在含有血清的培养基中时，细胞第一次结合到组织培养聚苯乙烯上是吸附到血清Vn表面的结果。血清Vn在表面上的吸附克服了血清成分的影响，后者往往会减少细胞的附着。
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.