Attachment and growth of anchorage-dependent cells on a novel, charged-surface microcarrier under serum-free conditions
Attachment and growth of anchorage-dependent cells on a novel, charged-surface microcarrier under serum-free conditions
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DOI：
10.1023/a:1008029715765
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发表时间：
1998-01-01
期刊：
影响因子：
2.2
通讯作者：
Hillegas, WJ
中科院分区：
文献类型：
--
作者：
Varani, J;Piel, F;Hillegas, WJ
本研究描述了一种新的微载体基板组成的可溶胀的，苯乙烯和二乙烯基苯的共聚物，衍生与三甲胺。共聚物三甲胺微载体支持许多不同细胞系- Madin达比牛肾、Madin-达比犬肾、Vero和Cos-7 -在无血清条件下生长，以及人二倍体成纤维细胞在含血清培养基中生长。细胞附着到共聚物三甲胺微载体上的速度与它们附着到其他带电表面微载体上的速度一样快（比它们附着到胶原蛋白包被的聚苯乙烯微载体上的速度更快），并且在附着后迅速扩散。所有检测的细胞在共聚物三甲胺微载体上生长至高密度。此外，细胞在暴露于胰蛋白酶/EDTA溶液后容易从表面释放。在这方面，共聚物三甲胺微载体不同于其他带电表面微载体。在该基质上生长的Madin-Darby牛肾细胞支持疫苗株传染性牛鼻气管炎病毒的生产，与在其他带电表面或胶原包被的微载体上一样容易。因此，共聚物三甲基胺微载体联合收割机在单一产品中结合了目前可用的带电表面和粘附肽包被的微载体的积极特征。病毒疫苗生产行业正在经历相当大的变化，制造商走向完整的，无动物产品的培养系统。这种新型基质应该在工业中得到应用，特别是在依赖于活细胞回收的过程中。
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.