Gümüşderelioğlu Menemşe, Türkoğlu Hilal
Chemical Engineering Department, Hacettepe University, Beytepe, Ankara, Turkey.
Biomaterials. 2002 Oct;23(19):3927-35. doi: 10.1016/s0142-9612(02)00128-x.
In this study, the development of a novel cell support material was purposed as due to the serum-free cultivation of tissue cells. This material was prepared by immobilizing RGD (Arg-Gly-Asp) sequence of cell-adhesion factor, fibronectin, and cell-growth factor, insulin, to the three-dimensional non-woven polyester fabric (briefly NWPF) discs that have been used successfully in our previous cell culture studies. At first these matrices were partially hydrolyzed and then the carboxyl groups were coupled with RGD or insulin in the presence of water-soluble carbodiimide. The effectiveness of immobilization process was checked with SEM, ATR-FTIR spectroscopy and swelling studies. The maximum amount of immobilized insulin was 6.96 micorgcm(-2) and it was obtained at 200 micorgml(-1) initial insulin concentration for 60 min immobilization period. The cell culture studies which were carried out with human skin fibroblasts (HS An1) showed that, percentage of adhesion on RGD modified NWPF discs is higher than that of other surfaces. i.e., unmodified discs, polystyrene Petri dishes and insulin-immobilized discs, in serum-free culture. According to the results of growth studies, highest cell yield was obtained in the case of insulin-modified discs.
在本研究中,由于组织细胞的无血清培养,旨在开发一种新型细胞支持材料。这种材料是通过将细胞粘附因子纤连蛋白的RGD(精氨酸-甘氨酸-天冬氨酸)序列和细胞生长因子胰岛素固定到三维非织造聚酯织物(简称NWPF)圆盘上制备而成的,该圆盘在我们之前的细胞培养研究中已成功使用。首先将这些基质部分水解,然后在水溶性碳二亚胺存在下将羧基与RGD或胰岛素偶联。通过扫描电子显微镜(SEM)、衰减全反射傅里叶变换红外光谱(ATR-FTIR)和溶胀研究来检查固定化过程的有效性。固定化胰岛素的最大量为6.96微克/平方厘米,在初始胰岛素浓度为200微克/毫升、固定化时间为60分钟时获得。用人皮肤成纤维细胞(HS An1)进行的细胞培养研究表明,在无血清培养中,RGD修饰的NWPF圆盘上的粘附百分比高于其他表面,即未修饰的圆盘、聚苯乙烯培养皿和胰岛素固定化圆盘。根据生长研究结果,胰岛素修饰的圆盘获得了最高的细胞产量。