Simultaneous flow cytometric analysis of cell surface markers and telomere length: analysis of human tonsilar B cells

被引:30
作者
Batliwalla, FM
Damle, RN
Metz, C
Chiorazzi, N
Gregersen, PK
机构
[1] N Shore Univ Hosp, NYU Sch Med, Dept Med, Div Biol & Human Genet, Manhasset, NY 11030 USA
[2] N Shore Univ Hosp, NYU Sch Med, Dept Med, Div Rheumatol Allergy & Clin Immunol, Manhasset, NY 11030 USA
[3] Picower Inst Med Res, Manhasset, NY 11030 USA
关键词
telomere length; immunophenotyping; B cells; flow cytometry;
D O I
10.1016/S0022-1759(00)00297-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Telomere Flow FISH is a recently developed method which allows the measurement of telomere length in purified subsets of cells using flow cytometry. However, the harsh conditions required for how FISH have precluded its use with conventional cell surface staining, thus limiting its utility for large scale clinical studies. We have now developed a method which permits simultaneous analysis of cell surface markers along with telomere length estimation by flow cytometry. This new assay employs the covalent crosslinking of monoclonal antibodies conjugated with a heat stable fluorochrome to the cell surface prior to flow FISH. Using this technique we have confirmed that human germinal center B cells (IgD(-)/CD38(+)) have dramatically longer telomeres than pre-germinal center founder B cells (IgD(+)/CD38(+)). This approach simplifies the analysis of complex cell populations and will facilitate widespread investigation of telomere length in health and disease states. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:103 / 109
页数:7
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