Rufer N, Dragowska W, Thornbury G, Roosnek E, Lansdorp P M
Terry Fox Laboratory, British Columbia Cancer Research Centre, Vancouver, Canada.
Nat Biotechnol. 1998 Aug;16(8):743-7. doi: 10.1038/nbt0898-743.
To measure the average length of telomere repeats at chromosome ends in individual cells we developed a flow cytometry method using fluorescence in situ hybridization (flow FISH) with labeled peptide nucleic acid (PNA) probes. Results of flow FISH measurements correlated with results of conventional telomere length measurements by Southern blot analysis (R = 0.9). Consistent differences in telomere length in CD8+ T-cell subsets were identified. Naive and memory CD4+ T lymphocytes in normal adults differed by around 2.5 kb in telomere length, in agreement with known replicative shortening of telomeres in lymphocytes in vivo. T-cell clones grown in vitro showed stabilization of telomere length after an initial decline and rare clones capable of growing beyond 100 population doublings showed variable telomere length. These results show that flow FISH can be used to measure specific nucleotide repeat sequences in single cells and indicate that the very large replicative potential of lymphocytes is only indirectly related to telomere length.
为了测量单个细胞染色体末端端粒重复序列的平均长度，我们开发了一种流式细胞术方法，该方法使用标记的肽核酸（PNA）探针进行荧光原位杂交（流式FISH）。流式FISH测量结果与通过Southern印迹分析进行的传统端粒长度测量结果相关（R = 0.9）。我们确定了CD8 + T细胞亚群中端粒长度的一致差异。正常成年人的初始和记忆性CD4 + T淋巴细胞的端粒长度相差约2.5 kb，这与体内淋巴细胞中端粒已知的复制性缩短一致。体外培养的T细胞克隆在最初下降后显示端粒长度稳定，极少数能够生长超过100次群体倍增的克隆显示出可变的端粒长度。这些结果表明，流式FISH可用于测量单细胞中的特定核苷酸重复序列，并表明淋巴细胞非常大的复制潜力仅与端粒长度间接相关。
