Gröhn P, Klöck G, Zimmermann U
Theodor-Boveri-Institut für Biowissenschaften, Würzburg, Germany.
Biotechniques. 1997 May;22(5):970-5. doi: 10.2144/97225rr06.
Several types of microcarriers suitable for large-scale cultivation of mammalian cells are commercially available. However, many of these carriers have disadvantages, e.g., the need for enzymatic digestion for cell harvesting, size limitations and insufficient biocompatibility. These limitations have been overcome by the development of collagen-coated Ba(2+)-alginate microcarriers. Ba(2+)-alginate microspheres, made with the air-jet droplet generator technique, were collagen-coated by incubation in a 0.5% collagen solution, with subsequent gelling of the collagen layer around the alginate microspheres. Human chang liver (CCL-13) and mouse fibroblast (L929) cell lines were cultivated in stationary, unstirred cultures as model systems. After a lag phase of nearly 24 h, the cells grew rapidly on these microcarriers and reached confluence after 3 days. The microcarrier cultures were stable for an additional 4-9 days and longer. Cells were harvested either by trypsinization or by dissolution of the alginate matrix using 5 mM EDTA. The main advantages of this new microcarrier system are that the preparation procedure is easy and can be accomplished on demand with standard laboratory equipment.
有几种适用于大规模培养哺乳动物细胞的微载体在市场上可以买到。然而，这些载体中的许多都有缺点，例如，细胞收获需要酶消化、尺寸限制和生物相容性不足。胶原包被的Ba(2+) - 海藻酸盐微载体的开发克服了这些限制。通过空气喷射液滴发生器技术制备的Ba(2+) - 海藻酸盐微球，在0.5%胶原溶液中孵育进行胶原包被，随后在海藻酸盐微球周围形成胶原层凝胶化。将人Chang肝(CCL - 13)和小鼠成纤维细胞(L929)细胞系作为模型系统，在静止、不搅拌的培养物中培养。经过近24小时的延迟期后，细胞在这些微载体上迅速生长，并在3天后达到汇合。微载体培养物在另外4 - 9天及更长时间内保持稳定。通过胰蛋白酶消化或使用5 mM EDTA溶解海藻酸盐基质来收获细胞。这种新型微载体系统的主要优点是制备过程简单，并且可以使用标准实验室设备按需完成。
