Cowan N J, Milstein C
Biochem J. 1972 Jun;128(2):445-54. doi: 10.1042/bj1280445.
An instrument is described that will maintain a population of mammalian cells at constant cell density while automatically monitoring the growth rate of the culture and the extent of precursor incorporation into a variety of cell products. The apparatus was used in an investigation of cyclic changes in the incorporation of labelled precursors into the DNA, RNA, total protein and myeloma protein synthesized in synchronous cultures of a mouse myeloma line. The incorporation of [(3)H]uridine into trichloroacetic acid-insoluble material reveals a slight periodicity, with maxima and minima corresponding to late S phase and the mitotic phases respectively. The incorporation of [(3)H]lysine into total intracellular protein also shows a slight oscillation, with maxima and minima occurring during the respective G2 and mitotic phases. Cyclical changes in the synthesis of serologically precipitable myeloma protein were found to vary somewhat according to the conditions used to synchronize the cells. In experiments conducted with 4.0mm-thymidine, maximal incorporation of label took place during S phase or early G2 phase. Experiments with 1.0mm-thymidine revealed a significantly less marked periodicity of myeloma protein synthesis.
本文描述了一种仪器,该仪器能够在自动监测培养物生长速率以及前体掺入多种细胞产物程度的同时,将哺乳动物细胞群体维持在恒定的细胞密度。该装置用于研究在小鼠骨髓瘤细胞系同步培养物中,标记前体掺入DNA、RNA、总蛋白和骨髓瘤蛋白过程中的周期性变化。将[³H]尿苷掺入三氯乙酸不溶性物质中显示出轻微的周期性,最大值和最小值分别对应于S期晚期和有丝分裂期。将[³H]赖氨酸掺入细胞内总蛋白中也显示出轻微的振荡,最大值和最小值分别出现在各自的G2期和有丝分裂期。发现血清学上可沉淀的骨髓瘤蛋白合成的周期性变化会根据用于使细胞同步化的条件而有所不同。在用4.0mM胸腺嘧啶核苷进行的实验中,标记物的最大掺入发生在S期或G2期早期。用1.0mM胸腺嘧啶核苷进行的实验显示,骨髓瘤蛋白合成的周期性明显不那么显著。