Benz C, Cadman E
Cancer Res. 1981 Jan;41(1):157-63.
The following parameters were evaluated at several points throughout unperturbed suspension culture growth of L1210 cells: cell volume; DNA histograms; the mean content of cellular DNA, RNA, and protein; ribonucleoside and deoxyribonucleoside triphosphate pools; phosphoribosyl pyrophosphate; and the incorporation of glycine into purine bases. The cell volume, the incorporation of glycine into purine bases, and the intracellular pools of phosphoribosyl pyrophosphate and dexoyribunucleotides began to decrease significantly during the midportion of logarithmic cell growth. However, there was no significant change in the DNA content per cell during culture growth. The RNA, protein content, and ribonucleotides all demonstrated a biphasic pattern with the highest values obtained during the midportion of logarithmic growth followed by rapid decline as the culture approached plateau growth. These intracellular fluctuations in de novo synthesis and precursor pools were correlated with the variable intracellular accumulation of three fluoropyrimidines (5-fluorouracil, 5-fluorouridine, and 5-fluorodeoxyuridine) and their active metabolites (5-fluorouridine triphosphate and 5-fluorodeoxyuridylate). These studies were performed to demonstrate that multiple biochemical alterations occur during logarithmically growing suspension cell cultures and could result in misleading conclusions of experiments with antimetabolites unless these factors are considered in the context of the performed studies.
在L1210细胞未受干扰的悬浮培养生长过程中的几个时间点评估了以下参数：细胞体积；DNA直方图；细胞DNA、RNA和蛋白质的平均含量；核糖核苷和脱氧核糖核苷三磷酸库；磷酸核糖焦磷酸；以及甘氨酸掺入嘌呤碱的情况。在对数期细胞生长的中期，细胞体积、甘氨酸掺入嘌呤碱的情况以及磷酸核糖焦磷酸和脱氧核糖核苷酸的细胞内库开始显著下降。然而，在培养生长过程中，每个细胞的DNA含量没有显著变化。RNA、蛋白质含量和核糖核苷酸均呈现双相模式，在对数生长中期达到最高值，随后随着培养接近平台期生长而迅速下降。这些从头合成和前体库中的细胞内波动与三种氟嘧啶（5-氟尿嘧啶、5-氟尿苷和5-氟脱氧尿苷）及其活性代谢物（5-氟尿苷三磷酸和5-氟脱氧尿苷酸）在细胞内的可变积累相关。进行这些研究是为了证明在对数生长的悬浮细胞培养过程中会发生多种生化改变，并且可能导致抗代谢物实验得出误导性结论，除非在进行的研究背景中考虑这些因素。
