Chinchar G D, Sinclair J H
J Cell Physiol. 1978 Sep;96(3):333-42. doi: 10.1002/jcp.1040960309.
Nutritional requirements of amphibian cells in culture were studied for the purpose of modifying a minimal medium in which frog cells could proliferate and which could be used for obtaining drug-resistant and auxotrophic variants. The serum, purine, CO2, and amino acid requirements for ICR 2A (a Rana pipiens haploid cell strain) have been investigated employing two different media: L-15, a nonbicarbonate, amino acid-buffered medium and Eagle's MEM, a bicarbonate-buffered medium. In this paper we present evidence to support the following conclusions: (1) With L-15 as the base medium, 10% fetal calf serum (FCS) supports optimal cell growth during exponential phase. Calf serum, whole, dialyzed, or heat-inactivated, cannot substitute for FCS and, in fact, is inhibitory. (2) Purines are required by ICR 2A cells only if grown in a nonbicarbonate-buffered medium, since the cells under these conditions cannot produce enough endogenous CO2 to support de novo purine synthesis. (3) In addition to the amino acids considered essential for mammalian cells in culture, ICR 2A cells depend upon exogenous asparagine. Glutamine and/or aspartic acid cannot replace the asparagine requirement. However, ICR 2A cells do utilized exogenous glutamine as an oxidative substrate.
为了改良一种能使蛙细胞增殖且可用于获得抗药和营养缺陷型变体的基础培养基，对培养中的两栖动物细胞的营养需求进行了研究。使用两种不同的培养基研究了ICR 2A（一种北美豹蛙单倍体细胞株）对血清、嘌呤、二氧化碳和氨基酸的需求：L-15，一种无碳酸氢盐、氨基酸缓冲的培养基，以及伊格尔氏MEM，一种碳酸氢盐缓冲的培养基。在本文中，我们提供证据支持以下结论：（1）以L-15作为基础培养基，10%胎牛血清（FCS）支持指数生长期的最佳细胞生长。小牛血清，无论是全血清、透析血清还是热灭活血清，都不能替代FCS，实际上还具有抑制作用。（2）只有在无碳酸氢盐缓冲的培养基中培养时，ICR 2A细胞才需要嘌呤，因为在这些条件下细胞无法产生足够的内源性二氧化碳来支持嘌呤的从头合成。（3）除了培养哺乳动物细胞所需的必需氨基酸外，ICR 2A细胞还依赖外源性天冬酰胺。谷氨酰胺和/或天冬氨酸不能替代对天冬酰胺的需求。然而，ICR 2A细胞确实利用外源性谷氨酰胺作为氧化底物。
