Aug 10, 2013

Investigation of telomere length dynamics in induced pluripotent stem cells using quantitative fluorescence in situ hybridization

Tissue & Cell
Masanori TeraiKaiyo Takubo

Abstract

Here we attempted to clarify telomere metabolism in parental cells and their derived clonal human induced pluripotent stem cells (iPSCs) at different passages using quantitative fluorescence in situ hybridization (Q-FISH). Our methodology involved estimation of the individual telomere lengths of chromosomal arms in individual cells within each clone in relation to telomere fluorescence units (TFUs) determined by Q-FISH. TFUs were very variable within the same metaphase spread and within the same cell. TFUs of the established iPSCs derived from human amnion (hAM933 iPSCs), expressed as mean values of the median TFUs of 20 karyotypes, were significantly longer than those of the parental cells, although the telomere extension rates varied quite significantly among the clones. Twenty metaphase spreads from hAM933 iPSCs demonstrated no chromosomal instability. The iPSCs established from fetal lung fibroblasts (MRC-5) did not exhibit telomere shortening and chromosomal instability as the number of passages increased. However, the telomeres of other iPSCs derived from MRC-5 became shorter as the number of passages increased, and one (5%) of 20 metaphase spreads showed chromosomal abnormalities including X trisomy at an early stage and...Continue Reading

  • References23
  • Citations4

Mentioned in this Paper

Metabolic Process, Cellular
Fluorescent in Situ Hybridization
Fetal Lung
Specimen Type - Fibroblasts
Telomere Lengthening
Protein Methylation
Chromosomes, Human, X
Telomere Shortening
Telomerase Catalytic Subunit
Transcription, Genetic

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