BIOSYNTHETIC LABELING WITH P-32 - RADIATION-DAMAGE TO MAMMALIAN-CELLS
BIOSYNTHETIC LABELING WITH P-32 - RADIATION-DAMAGE TO MAMMALIAN-CELLS
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DOI：
10.1016/0003-2697(85)90125-3
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发表时间：
1985-01-01
影响因子：
2.9
通讯作者：
BURGESS, AW
中科院分区：
文献类型：
--
作者：
COOPER, PC;BURGESS, AW
理论计算表明，使用典型浓度的32 P（1 mCi/ml）对细胞进行生物合成放射性标记导致细胞吸收高辐射剂量（200-500 rad/h）。随后对小鼠骨髓单核细胞白血病细胞系（WEHI-3B（D+））的研究表明，在短暂（不到1小时）暴露于1 mCi/ml的32 P时，复制能力显著丧失。在同位素剂量小于100拉德时发现细胞复制能力完全丧失（即，100 μ Ci/ml持续5小时）。采用辐射敏感性较低的前B细胞系（18.81）的实验显示，在与WEHI-3B（D+）细胞系相同的条件下，在与32 P孵育期间发生活力显著丧失。利用衰变批次的32 P和从细胞中物理分离同位素溶液的对照实验证实，细胞毒性是由辐射发射引起的，而不是同位素溶液中存在毒性组分。计算了~（59）Fe、~（33）P、~（35）S和~（14）C标记细胞的辐射吸收剂量。虽然32 P可以吸收大量的辐射，但它的放射性比其他同位素要大得多。计算结果表明，合理地选择容器的几何形状可以降低32 P溶液的辐射吸收剂量。特别是在毛细管（直径小于1 mm）中对细胞进行生物合成放射性标记可将吸收率降低至小于悬浮在培养皿或离心管中的细胞所接受剂量的1/10。
Theoretical calculations showed that biosynthetic radiolabeling of cells using typical concentrations of 32P (1 mCi/ml) resulted in high radiation doses (200-500 rad/h) being absorbed by the cells. Subsequent investigations with a mouse myelomonocytic leukemia cell line (WEHI-3B(D+)) showed significant loss of replicative ability during brief (less than 1 h) exposures to 1 mCi/ml of 32P. Complete loss of cell replicative ability was found with isotopic doses less than 100 rad (i.e., 100 .mu.Ci/ml for 5 h). Experiments employing a less radiosensitive pre-B-cell line (18.81) revealed that significant loss of viability occurred during incubation with 32P under identical conditions to those employed for the WEHI-3B(D+) cell line. Control experiments utilizing decayed batches of 32P and physical separation of the isotope solution from the cells confirmed that the cytotoxicity was caused by radiation emission rather than the presence of toxic components in the isotopic solution. The radiation doses absorbed by cells biosynthetically labeled with 59Fe, 33P, 35S and 14C were calculated. Although significant levels of radiation can be absorbed 32P was considerably more radiotoxic than the other isotopes. The results of calculations indicated that the judicious choice of container geometry could reduce the absorbed radiation dose from 32P solutions. In particular the biosynthetic radiolabeling of cells in capillary tubes (diameter less than 1 mm) can reduce the absorbed rate to less than 1/10 of the dose received by cells suspended in Petri dishes or centrifuge tubes.