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Investigation of telomere length dynamics in induced pluripotent stem cells using quantitative fluorescence in situ hybridization
- Source :
- Tissuecell. 45(6)
- Publication Year :
- 2013
-
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 all 20 showed abnormalities including X and 12 trisomies at the late stage.
- Subjects :
- Chromosomes, Human, X
Chromosomes, Human, Pair 12
Induced Pluripotent Stem Cells
Clone (cell biology)
Q-FISH
Chromosome
Telomere Homeostasis
Karyotype
Trisomy
Cell Biology
General Medicine
Biology
Telomere
Molecular biology
Chromosome instability
Karyotyping
Humans
Induced pluripotent stem cell
Metaphase
Telomerase
In Situ Hybridization, Fluorescence
Developmental Biology
Subjects
Details
- ISSN :
- 15323072
- Volume :
- 45
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Tissuecell
- Accession number :
- edsair.doi.dedup.....02a879101e83d231dd63319e376950ce