Telomere length measurements in leukocyte subsets by automated multicolor flow-FISH

被引:81
作者
Baerlocher, GM
Lansdorp, PM
机构
[1] British Columbia Canc Agcy, Terry Fox Lab, Vancouver, BC V5Z 1L3, Canada
[2] Univ British Columbia, Dept Med, Vancouver, BC V5Z 1M9, Canada
来源
CYTOMETRY PART A | 2003年 / 55A卷 / 01期
关键词
telomere; telomere length; multicolor flow; fluorescence in situ hybridization; automation;
D O I
10.1002/cyto.a.10064
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Background: Telomeres are essential protein-DNA structures at the end of chromosomes which are implicated in genome stability and cell replication. The average length of telomere rep eats can be measured by in situ hybridization and flow cytometry [flow-FISH]. Such telomere length values reflect telomere shortening (resulting from cell divisions, oxidative damage and other causes) and telomere elongation (mainly resulting from telomerase activity) of the chromosome-specific telomere length inherited in the gametes. Here we report improvements in flow-FISH methodology that enable measurements of telomere length in subsets of human nucleated blood cells. Methods and Results: In order to measure the telomere length in granulocytes, naive T cells, memory T cells, B cells and natural killer (NK)/NKT cells within a blood sample, we combined flow-FISH with antibody-staining (Multicolor flow-FISH). Most steps in the staining protocol were automated using a 96-well microdispenser device. The minimum detectable difference in telomere length and the reproducibility of the method are in the range of 0.2-0.5 kb and measurements can be made with as few as a thousand cells. Conclusions: Automated multicolor flow-FISH will greatly facilitate studies of telomere length regulation in subsets of nucleated blood cells, especially when only few cells are available and when differences in telomere length are small. (C) 2003 Wiley-Liss, Inc.
引用
收藏
页码:1 / 6
页数:6
相关论文
共 25 条
  • [1] Telomere length measurement by fluorescence in situ hybridization and flow cytometry: Tips and pitfalls
    Baerlocher, GM
    Mak, J
    Tien, T
    Lansdorp, PM
    [J]. CYTOMETRY, 2002, 47 (02): : 89 - 99
  • [2] Simultaneous flow cytometric analysis of cell surface markers and telomere length: analysis of human tonsilar B cells
    Batliwalla, FM
    Damle, RN
    Metz, C
    Chiorazzi, N
    Gregersen, PK
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 2001, 247 (1-2) : 103 - 109
  • [3] Telomere states and cell fates
    Blackburn, EH
    [J]. NATURE, 2000, 408 (6808) : 53 - 56
  • [4] Extension of life-span by introduction of telomerase into normal human cells
    Bodnar, AG
    Ouellette, M
    Frolkis, M
    Holt, SE
    Chiu, CP
    Morin, GB
    Harley, CB
    Shay, JW
    Lichtsteiner, S
    Wright, WE
    [J]. SCIENCE, 1998, 279 (5349) : 349 - 352
  • [5] Analysis of telomeric repeats and telomerase activity in human colon carcinoma cells with gene amplification
    Bolzán, AD
    Páez, GL
    Bianchi, MS
    Bianchi, NO
    [J]. CANCER GENETICS AND CYTOGENETICS, 2000, 120 (02) : 166 - 170
  • [6] Cellular senescence as a tumor-suppressor mechanism
    Campisi, J
    [J]. TRENDS IN CELL BIOLOGY, 2001, 11 (11) : S27 - S31
  • [7] Telomerase in the human organism
    Collins, K
    Mitchell, JR
    [J]. ONCOGENE, 2002, 21 (04) : 564 - 579
  • [8] Telomere dynamics, end-to-end fusions and telomerase activation during the human fibroblast immortalization process
    Ducray, C
    Pommier, JP
    Martins, L
    Boussin, FD
    Sabatier, L
    [J]. ONCOGENE, 1999, 18 (29) : 4211 - 4223
  • [9] IDENTIFICATION OF A SPECIFIC TELOMERE TERMINAL TRANSFERASE-ACTIVITY IN TETRAHYMENA EXTRACTS
    GREIDER, CW
    BLACKBURN, EH
    [J]. CELL, 1985, 43 (02) : 405 - 413
  • [10] TELOMERES SHORTEN DURING AGING OF HUMAN FIBROBLASTS
    HARLEY, CB
    FUTCHER, AB
    GREIDER, CW
    [J]. NATURE, 1990, 345 (6274) : 458 - 460