Department of Pathology, The University of Michigan, 1301 Catherine Road, P.O. Box 0602, Ann Arbor, MI, 48109, U.S.A.,
Cytotechnology. 1998 Nov;28(1-3):101-9. doi: 10.1023/A:1008029715765.
The present study describes a novel microcarrier substrate consisting of a swellable, copolymer of styrene and divinylbenzene, derivatized with trimethylamine. The co-polymer trimethylamine microcarriers support the growth of a number of different cell lines - Madin Darby Bovine Kidney, Madin-Darby Canine Kidney, Vero and Cos-7 - under serum-free conditions, and human diploid fibroblasts in serum-containing medium. Cells attach to the co- polymer trimethylamine microcarriers as rapidly as they attach to other charged-surface microcarriers (faster than they attach to collagen-coated polystyrene microcarriers) and spread rapidly after attachment. All of the cells examined grow to high density on the co- polymer trimethylamine microcarriers. Furthermore, cells are readily released from the surface after exposure to a solution of trypsin/EDTA. In this respect, the co-polymer trimethylamine microcarriers are different from other charged-surface microcarriers. Madin-Darby Bovine Kidney cells grown on this substrate support production of vaccine strain infectious bovine rhinotracheitis virus as readily as on other charged-surface or collagen-coated microcarriers. Thus, the co-polymer trimethylamine microcarriers combine the positive characteristics of the currently available charged-surface and adhesion-peptide coated microcarriers in a single product. The viral vaccine production industry is undergoing considerable change as manufacturers move toward complete, animal product-free culture systems. This novel substrate should find application in the industry, especially in processes which depend on viable cell recovery.
本研究描述了一种新型的微载体基质，由可溶胀的苯乙烯和二乙烯基苯的共聚物衍生而来，并用三甲胺进行了修饰。共聚三甲胺微载体在无血清条件下支持多种不同细胞系（马-达二氏牛肾细胞、马-达二氏犬肾细胞、vero 细胞和 Cos-7 细胞）的生长，并在含血清的培养基中支持人类二倍体成纤维细胞的生长。细胞附着在共聚物三甲胺微载体上的速度与其他带电表面微载体（比附着在胶原涂层聚苯乙烯微载体上的速度更快）一样快，并且在附着后迅速扩散。所有检查的细胞在共聚物三甲胺微载体上生长到高密度。此外，细胞在用胰蛋白酶/EDTA 溶液处理后很容易从表面释放。在这方面，共聚物三甲胺微载体与其他带电表面微载体不同。在这种基质上生长的马-达二氏牛肾细胞支持疫苗株传染性牛鼻气管炎病毒的生产，与其他带电表面或胶原涂层微载体一样容易。因此，共聚三甲胺微载体将目前可用的带电表面和粘附肽涂层微载体的优点结合在一个单一的产品中。随着制造商向完全无动物产品的培养系统转变，病毒疫苗生产行业正在发生重大变化。这种新型基质应该在该行业找到应用，特别是在依赖活细胞回收的工艺中。
