Quantifying telomere lengths of human individual chromosome arms by centromere-calibrated fluorescence in situ hybridisation and digital imaging

Perner, Sven; Brüderlein, Silke; Hasel, Cornelia; Waibel, I.; Holdenried, A.; Ciloglu, Neslisah; Chopurian, Heiko; Nielsén, Karin; Plesch, Andreas; Högel, Josef; Möller, Peter

doi:10.1016/s0344-0338(04)80508-9

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“…The reliability and limitations of PNA-FISH-based telomere length measurements using the same equipment which was used here, have recently been discussed in detail (Perner et al, 2003). Facing the interchromosomal (Londono-Vallejo et al, 2001), the intercellular, and the interindividual (Lansdorp et al, 1996;Perner et al, 2003) variation of telomere lengths which could also be confirmed in the present study, differences of telomere lengths should be rather clear to be statistically significant. For instance, the comparison of p-and q-arm telomeres of the normal chromosomes of the cell line examined here did not yield the differences which were found in mouse chromosomes (Modino and Slijepcevic, 2002).…”

Section: Discussionsupporting

confidence: 63%

“…All figures, therefore, are given as percentage of 2cen fluorescence intensity. The technical details of telomere analysis by this system have been described previously by others (Perner et al, 2003). Because of the limitations of the evaluation system, normal chromosomes and marker chromosomes had to be karyotyped and evaluated separately from identical metaphases.…”

Section: Microscopic Detection and Image Analysismentioning

confidence: 99%

“…We have examined the telomeres of a karyotypically rather well characterized Tcell acute lymphoblastic leukemia (T-ALL) cell line (CCRF-CEM) with several defined markers (Weise et al, 2002) to compare the telomere lengths of these markers with those of their chromosomes of origin. Peptide nucleic acid (PNA) probes were applied for FISH detection of telomeres and quantitative estimation of their relative lengths, as these probes have been shown to be highly specific and most efficient for this purpose (Lansdorp et al, 1996;Martens et al, 1998;Perner et al, 2003). As a support to correct assignment of the end structures of the marker chromosomes several additional DNA probes were included in this study (i.e.…”

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confidence: 99%

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FISH studies on the telomeric regions of the T-cell acute lymphoblastic leukemia cell line CCRF-CEM

Wick

Kirsch²,

Rauch³

et al. 2005

Cytogenet Genome Res

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So far, the problem of an influence of translocations on the telomeres of the involved chromosomes has not been addressed yet in human cells. Therefore, the telomeres of a karyotypically rather well characterized T-cell acute lymphoblastic leukemia (T-ALL) cell line (CCRF-CEM) with several marker chromosomes were examined using peptide nucleic acid (PNA) telomere FISH probes to compare the telomere length of these markers with that of the chromosome arms of their origin. In addition, chromosome libraries, centromeric probes, and subtelomeric DNA probes were used to further define the marker chromosomes. Two markers could be newly defined and a concise karyotype of the cell line could be obtained by these detailed examinations: 42–47,X,–X,del(5) (q35?),t(5;15)(q14;q13.2),t(8;9)(p11;p24),del(9)(:p13→qter)/inv(9)(pter→p12::q21→p12::q21→qter),+13,+20,+der(22)(p+ [HSR?])[cp]. The relative telomere length of all chromosomes showed considerable interchromosomal, intercellular, and inter-passage variation. However, it could be shown, that in four different passages of the examined cell line the observed differences between relative telomere lengths of the markers and the chromosomes of their origin, with two exceptions (short arms of del/inv9 and der22), were not significant. On the other hand, because of its mentioned variability, telomere length alone is not sufficient to reliably define the derivation of markers.

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