title: [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].
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.
authors.affiliation: Department of Hematology, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
authors.forename: Tian-Jiao
authors.initials: TJ
authors.lastname: Guo
authors.forename: Wan-Ling
authors.initials: WL
authors.lastname: Sun
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Xiao-Cai
authors.initials: XC
authors.lastname: Ma
authors.forename: Cong-Yan
authors.initials: CY
authors.lastname: Liu
authors.forename: Jing-Juan
authors.initials: JJ
authors.lastname: He
authors.forename: Juan
authors.initials: J
authors.lastname: Xu
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D018952:Antigens, CD34 / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology; D004247:DNA; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009190:Myelodysplastic Syndromes / Q000235:genetics*; D016615:Telomere / Q000235:genetics*
chemical_list: D018952:Antigens, CD34; D004247:DNA