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Detection of apoptotic cells by selective precipitation of [3H]thymidine-labelled DNA.通过[3H]胸腺嘧啶核苷标记的DNA的选择性沉淀检测凋亡细胞。
Clin Diagn Lab Immunol. 1996 Jan;3(1):1-4. doi: 10.1128/cdli.3.1.1-4.1996.
2
A new flow cytometric method for discrimination of apoptotic cells and detection of their cell cycle specificity through staining of F-actin and DNA.一种通过F-肌动蛋白和DNA染色来区分凋亡细胞并检测其细胞周期特异性的新型流式细胞术方法。
Cytometry. 1995 Jun 1;20(2):162-71. doi: 10.1002/cyto.990200209.
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The proliferation response of rat liver parenchymal cells after partial hepatectomy. A methodological study comparing flow cytometry of nuclear DNA content and in vivo and in vitro uptake of thymidine.部分肝切除术后大鼠肝实质细胞的增殖反应。一项比较核DNA含量流式细胞术以及胸苷体内外摄取的方法学研究。
Cell Tissue Kinet. 1982 Sep;15(5):521-8. doi: 10.1111/j.1365-2184.1982.tb01574.x.
4
Effect of curcumin on cell cycle progression and apoptosis in vascular smooth muscle cells.姜黄素对血管平滑肌细胞细胞周期进程及凋亡的影响
Br J Pharmacol. 1998 Jul;124(6):1029-40. doi: 10.1038/sj.bjp.0701914.
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Analysis of apoptosis by propidium iodide staining and flow cytometry.通过碘化丙啶染色和流式细胞术分析细胞凋亡。
Nat Protoc. 2006;1(3):1458-61. doi: 10.1038/nprot.2006.238.
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Adaptation of a simple flow cytometric assay to identify different stages during apoptosis.
Cytometry. 1996 Oct 1;25(2):173-81. doi: 10.1002/(SICI)1097-0320(19961001)25:2<173::AID-CYTO6>3.0.CO;2-I.
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Flow cytometric detection of apoptosis: comparison of the assays of in situ DNA degradation and chromatin changes.流式细胞术检测细胞凋亡：原位DNA降解和染色质变化检测方法的比较。
Cytometry. 1994 Mar 1;15(3):237-44. doi: 10.1002/cyto.990150309.
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Discrepancies between flow cytometric analysis and [3H]thymidine incorporation in stimulated lymphocytes.流式细胞术分析与[3H]胸腺嘧啶核苷掺入刺激淋巴细胞结果之间的差异。
Biochim Biophys Acta. 1983 Jun 2;762(3):452-7. doi: 10.1016/0167-4889(83)90011-3.
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A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.一种通过碘化丙啶染色和流式细胞术检测胸腺细胞凋亡的快速简便方法。
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Detection of apoptotic cells using propidium iodide staining.使用碘化丙啶染色检测凋亡细胞。
Cold Spring Harb Protoc. 2014 Nov 3;2014(11):1202-6. doi: 10.1101/pdb.prot082545.

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Stat1-dependent, p53-independent expression of p21(waf1) modulates oxysterol-induced apoptosis.p21（waf1）的Stat1依赖性、p53非依赖性表达调节氧化甾醇诱导的细胞凋亡。
Mol Cell Biol. 2002 Apr;22(7):1981-92. doi: 10.1128/MCB.22.7.1981-1992.2002.
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CD4+-T-cell counts, spontaneous apoptosis, and Fas expression in peripheral blood mononuclear cells obtained from human immunodeficiency virus type 1-infected subjects.从1型人类免疫缺陷病毒感染受试者获得的外周血单个核细胞中的CD4 + T细胞计数、自发凋亡和Fas表达。
Clin Diagn Lab Immunol. 1997 Nov;4(6):736-41. doi: 10.1128/cdli.4.6.736-741.1997.

本文引用的文献

1
Apoptosis.细胞凋亡
Immunol Today. 1993 Mar;14(3):126-30. doi: 10.1016/0167-5699(93)90214-6.
2
Simultaneous determination of cell surface antigens and apoptosis.细胞表面抗原与细胞凋亡的同步检测
Trends Genet. 1994 Feb;10(2):41-2. doi: 10.1016/0168-9525(94)90140-6.
3
Calpain activation in apoptosis.细胞凋亡中的钙蛋白酶激活
J Cell Physiol. 1994 May;159(2):229-37. doi: 10.1002/jcp.1041590206.
4
Use of a flow cytometric assay to quantitate apoptosis in human lymphocytes.使用流式细胞术检测法定量人类淋巴细胞中的细胞凋亡。
Clin Immunol Immunopathol. 1994 Apr;71(1):14-8. doi: 10.1006/clin.1994.1045.
5
Accelerated apoptosis in peripheral blood mononuclear cells (PBMCs) from human immunodeficiency virus type-1 infected patients and in CD4 cross-linked PBMCs from normal individuals.来自人类免疫缺陷病毒1型感染患者的外周血单个核细胞（PBMC）以及来自正常个体的CD4交联PBMC中凋亡加速。
Blood. 1993 Dec 1;82(11):3392-400.
6
HIV type 1 Tat protein induces apoptosis and death in Jurkat cells.1型人类免疫缺陷病毒反式激活蛋白诱导Jurkat细胞凋亡和死亡。
AIDS Res Hum Retroviruses. 1995 Apr;11(4):443-50. doi: 10.1089/aid.1995.11.443.
7
The end of the (cell) line: methods for the study of apoptosis in vitro.
Methods Cell Biol. 1995;46:153-85. doi: 10.1016/s0091-679x(08)61929-9.
8
Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis.细胞凋亡中的染色质裂解：与浓缩染色质形态的关联及对大分子合成的依赖性。
J Pathol. 1984 Jan;142(1):67-77. doi: 10.1002/path.1711420112.
9
Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis.内源性核酸内切酶诱导的DNA片段化：细胞介导的细胞溶解中的早期事件。
Proc Natl Acad Sci U S A. 1983 Oct;80(20):6361-5. doi: 10.1073/pnas.80.20.6361.
10
Selective extraction of polyoma DNA from infected mouse cell cultures.从受感染的小鼠细胞培养物中选择性提取多瘤病毒DNA。
J Mol Biol. 1967 Jun 14;26(2):365-9. doi: 10.1016/0022-2836(67)90307-5.

通过[3H]胸腺嘧啶核苷标记的DNA的选择性沉淀检测凋亡细胞。

Detection of apoptotic cells by selective precipitation of [3H]thymidine-labelled DNA.

作者信息

Patki A H, Lederman M M

机构信息

Department of Medicine, Case Western Reserve University School of Medicine, University Hospitals of Cleveland, Ohio 44106, USA.

出版信息

Clin Diagn Lab Immunol. 1996 Jan;3(1):1-4. doi: 10.1128/cdli.3.1.1-4.1996.

DOI:10.1128/cdli.3.1.1-4.1996
PMID:8770495
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC170238/
Abstract

Apoptosis is characterized by systematic fragmentation of high-molecular-weight DNA into oligonucleosome fragments. A sensitive method for detection of apoptotic cells involving [3H]thymidine-labelled DNA is presented. Cells from mid-log-phase cultures were labelled with [3H]thymidine for 15 to 18 h and then exposed to gamma irradiation to induce apoptosis. A modified Hirt method was used to separate low-molecular-weight DNA from high-molecular-weight DNA. The percentage of fragmented DNA and high-molecular-weight DNA were measured by scintillation spectrometry. This method was compared with an established flow cytometric method for detection of apoptotic cells by using propidium iodide staining of DNA. We observed a good correlation between these two methods in detecting apoptosis. Hence, expensive flow cytometric assays for detection of apoptosis in dividing cells may be replaceable by a method involving [3H]thymidine labelling of DNA and separation of low-molecular-weight DNA from high-molecular-weight DNA by precipitation.

摘要

细胞凋亡的特征是高分子量DNA系统性断裂成寡核小体片段。本文介绍了一种检测凋亡细胞的灵敏方法，该方法涉及用[³H]胸腺嘧啶核苷标记DNA。对数中期培养的细胞用[³H]胸腺嘧啶核苷标记15至18小时，然后暴露于γ射线照射以诱导细胞凋亡。采用改良的赫特方法将低分子量DNA与高分子量DNA分离。通过闪烁光谱法测量断裂DNA和高分子量DNA的百分比。通过用碘化丙啶对DNA染色，将该方法与一种既定的流式细胞术检测凋亡细胞的方法进行比较。我们观察到这两种方法在检测细胞凋亡方面具有良好的相关性。因此，用于检测分裂细胞中细胞凋亡的昂贵流式细胞术检测方法可能可被一种涉及用[³H]胸腺嘧啶核苷标记DNA并通过沉淀从高分子量DNA中分离低分子量DNA的方法所替代。