Wunderlich K, Knorr M, Dartsch P C
Universitäts-Augenklinik, Tübingen, Germany.
Graefes Arch Clin Exp Ophthalmol. 1994 Jun;232(6):355-60. doi: 10.1007/BF00175987.
Mammalian cells in culture have individual nutritional requirements which are mainly fulfilled by the addition of fetal calf serum to the basic culture medium. Since many of the serum components are as yet poorly understood or even completely unknown, a number of difficulties arise in the evaluation of the effect of exogenously added factors or drugs on the growth of the cells. This paper presents data demonstrating the successful adaptation, routine cultivation and cryopreservation of bovine lens epithelial cells (BLEC) under serum-free culture conditions by use of the commercially available serum substitutes BMS, BM-86 Wissler and Ultroser G. Moreover, a culture medium especially designed for cell quiescence, named LR-1, is presented. Cells cultivated in culture medium containing 10% fetal calf serum served as controls. While BM-86 Wissler caused significantly reduced growth rates within 2 days and, finally, cell death after 12 days of incubation, the use of BMS resulted in growth rates which did not differ from the corresponding controls. Ultroser G resulted in a significant increase of proliferative activity of BLEC. LR-1 medium caused cell quiescence and kept the cells alive for a number of days. Thus, LR-1 allowed evaluation of the response of the cells to a mitogenic mixture from bovine brain mainly containing endothelial cell growth factor. The results demonstrate that cultivation of BLEC is possible under serum-free culture conditions. Moreover, the medium LR-1, which causes cell quiescence, is useful for the evaluation of growth factor-induced effects in vitro.
培养中的哺乳动物细胞有各自的营养需求,主要通过向基础培养基中添加胎牛血清来满足。由于许多血清成分尚未被充分了解甚至完全未知,在评估外源添加因子或药物对细胞生长的影响时会出现一些困难。本文展示了利用市售血清替代品BMS、BM - 86 Wissler和Ultroser G在无血清培养条件下成功适应、常规培养和冻存牛晶状体上皮细胞(BLEC)的数据。此外,还介绍了一种专门为细胞静止设计的培养基LR - 1。在含10%胎牛血清的培养基中培养的细胞用作对照。虽然BM - 86 Wissler在2天内导致生长速率显著降低,最终在培养12天后细胞死亡,但使用BMS导致的生长速率与相应对照无差异。Ultroser G导致BLEC的增殖活性显著增加。LR - 1培养基使细胞静止并使细胞存活了若干天。因此,LR - 1允许评估细胞对主要含内皮细胞生长因子的牛脑促有丝分裂混合物的反应。结果表明在无血清培养条件下培养BLEC是可行的。此外,能使细胞静止的LR - 1培养基对于评估体外生长因子诱导的效应很有用。