Application of flow cytometry-fluorescence in situ hybridization in the measurement of relative telomere length of bone marrow CD34(+) cells in patients with myelodysplastic syndrome.
10.7534/j.issn.1009-2137.2013.05.022
- Author:
Tian-Jiao GUO
1
;
Wan-Ling SUN
;
Wei ZHANG
;
Xiao-Cai MA
;
Cong-Yan LIU
;
Jing-Juan HE
;
Juan XU
Author Information
1. Department of Hematology, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
- Publication Type:Journal Article
- MeSH:
Aged;
Aged, 80 and over;
Antigens, CD34;
immunology;
Bone Marrow Cells;
cytology;
immunology;
DNA;
Female;
Flow Cytometry;
methods;
Humans;
In Situ Hybridization, Fluorescence;
methods;
Male;
Middle Aged;
Myelodysplastic Syndromes;
genetics;
Telomere;
genetics
- From:
Journal of Experimental Hematology
2013;21(5):1195-1199
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the feasibility of flow cytometry-fluorescence in situ hybridization (Flow-FISH) in measuring the telomere length of bone marrow cell subgroups in patients with myelodysplastic syndrome (MDS). Seven newly diagnosed patients with low-risk MDS and seven nutritional anemia patients who were matched with age and sex, were enrolled in this study. Heparinized bone marrow were sampled. Taking Molt-4 cell line as internal control cells, leukocytes isolated from whole bone marrow were labeled with CD34-Alexa Fluork ® 647, then denatured by high temperature and hybridized with FITC-conjugated telomere probe. The DNA was counterstained and the relative telomere length (RTL) of nucleated cells and CD34(+) cells in bone marrow were measured by four-color flow cytometry. The results showed that CD34(+) cells could be gated for the measurement of RTL in both groups, undergoing the denaturation and hybridization. Primary analysis indicated that the RTL of bone marrow CD34(+) cells in MDS patients was significantly shorter than that of bone marrow nucleated cells (P = 0.001), and the RTL of both CD34(+) cells and nucleated cells in bone marrow of MDS patients were significantly shorter than that of control group (P = 0.020, 0.002). It is concluded that the application of Flow-FISH in the measurement of RTL of certain cell subgroup is feasible by labeling the cell with thermostable fluorescence-conjugated antibody, and this technique is worthy to be investigated further.
