Jacobson B S, Ryan U S
Tissue Cell. 1982;14(1):69-83. doi: 10.1016/0040-8166(82)90008-8.
A new cell culture microcarrier that can be covalently bonded by cell attachment proteins and can be thin-sectioned for electron microscopy was synthesized. It was easily made by sulfonating cross-linked polystyrene beads for a negative surface charge followed by covalent attachment of polyethylenimine for a positive charge. Cell attachment proteins, e.g. collagen was covalently bonded directly to the microcarrier using a carbodiimide or after activating the microcarrier surface with glutaraldehyde. HeLa-S3 cells attached, spread and grew to confluence more efficiently on the positive microcarriers and those coated with collagen than on the negative ones. Endothelial cells, grew best on those with a negative surface charge. The nature of the microcarrier surface was not the only aspect involved in cell adhesion but also the type of serum proteins absorbed. Qualitatively different proteins coated the microcarriers depending upon whether the carrier was negative, positive or coated with collagen. Comparison of various types of available microcarriers indicated that the modified cross-linked polystyrene beads used here were best for transmission and scanning electron microscopy. Endothelial cells grown on the microcarriers had the same ultrastructure as cells grown in monolayers in culture dishes. Of a variety of microcarriers tested the modified cross-linked polystyrene beads were the only ones that could be used for both ultrastructural and biochemical techniques.
合成了一种新型细胞培养微载体，它可通过细胞黏附蛋白共价结合，并且可以制成薄片用于电子显微镜观察。它很容易制备，先将交联聚苯乙烯珠磺化以产生负表面电荷，然后共价连接聚乙烯亚胺以产生正电荷。细胞黏附蛋白，如胶原蛋白，可使用碳二亚胺直接共价结合到微载体上，或者在用戊二醛活化微载体表面后进行结合。HeLa-S3细胞在带正电荷的微载体和包被有胶原蛋白的微载体上比在带负电荷的微载体上更有效地附着、铺展并生长至汇合。内皮细胞在表面带负电荷的微载体上生长最佳。微载体表面的性质不是细胞黏附中涉及的唯一方面，吸附的血清蛋白类型也有影响。根据微载体是带负电荷、正电荷还是包被有胶原蛋白，微载体上包被的蛋白质在性质上有所不同。对各种可用微载体的比较表明，这里使用的改性交联聚苯乙烯珠最适合用于透射电子显微镜和扫描电子显微镜观察。在微载体上生长的内皮细胞具有与在培养皿中单层生长的细胞相同的超微结构。在测试的各种微载体中，改性交联聚苯乙烯珠是唯一可用于超微结构和生化技术的微载体。
