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胸苷和溴脱氧尿苷的示踪剂量及有效时间:溴脱氧尿苷在细胞动力学研究中的应用

Tracer dose and availability time of thymidine and bromodeoxyuridine: application of bromodeoxyuridine in cell kinetic studies.

作者信息

Böswald M, Harasim S, Maurer-Schultze B

机构信息

Institut für Medizinische Strahlenkunde, University of Würzburg, F.R. Germany.

出版信息

Cell Tissue Kinet. 1990 May;23(3):169-81. doi: 10.1111/j.1365-2184.1990.tb01113.x.

Abstract

The present experiments with [14C]-thymidine (TdR) and [3H]-bromodeoxyuridine (BrdU) using mouse jejunal crypt cells show that the upper limit of the tracer dose of TdR is about 0.5 microgram g body weight-1 and that of BrdU is about 5.0 micrograms g body weight-1. Applying these doses, the proportions of the endogenous DNA synthesis attributed to the exogenous DNA precursor are 2% and 9% respectively. For [3H]-TdR doses commonly used in cell kinetic studies this proportion is only 0.1-1.0%, a negligible quantity that does not influence the endogenous DNA synthesis. The maximum availability time of tracer doses of TdR as well as BrdU is 40 to 60 min, the majority of the precursors being incorporated after 20 min. The availability time is the same for TdR doses exceeding the tracer dose by a factor of 80, whereas it is prolonged in the case of BrdU doses exceeding the tracer dose by a factor of 50. BrdU is suitable to replace radioactively labelled TdR in short term cell kinetic studies, i.e. determination of the labelling index or of the S phase duration by double labelling. However, more studies are needed to elucidate how far BrdU can replace TdR in long term studies as shown by differences between the fraction of labelled mitoses (FLM) curves of a human renal cell carcinoma measured with BrdU and [3H]-TdR.

摘要

目前使用小鼠空肠隐窝细胞进行的[14C] - 胸腺嘧啶核苷(TdR)和[3H] - 溴脱氧尿苷(BrdU)实验表明,TdR示踪剂量的上限约为0.5微克/克体重,BrdU的上限约为5.0微克/克体重。应用这些剂量时,归因于外源性DNA前体的内源性DNA合成比例分别为2%和9%。对于细胞动力学研究中常用的[3H] - TdR剂量,该比例仅为0.1 - 1.0%,这是一个可忽略不计的量,不会影响内源性DNA合成。TdR以及BrdU示踪剂量的最大可用时间为40至60分钟,大多数前体在20分钟后被掺入。对于超过示踪剂量80倍的TdR剂量,可用时间相同,而对于超过示踪剂量50倍的BrdU剂量,可用时间会延长。在短期细胞动力学研究中,即通过双重标记测定标记指数或S期持续时间时,BrdU适合替代放射性标记的TdR。然而,正如用BrdU和[3H] - TdR测量的人肾细胞癌标记有丝分裂分数(FLM)曲线之间的差异所示,需要更多研究来阐明在长期研究中BrdU能在多大程度上替代TdR。

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