Investigation of telomere length dynamics in induced pluripotent stem cells using quantitative fluorescence in situ hybridization.
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.
Research Team for Geriatric Pathology, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0015, Japan; Department of Judotherapy, Faculty of Health Sciences, Tokyo Ariake University of Medical and Health Sciences, Tokyo 135-0063, Japan. Electronic address: terai@tau.ac.jp.
Masanori
M
Terai
Naotaka
N
Izumiyama-Shimomura
Junko
J
Aida
Naoshi
N
Ishikawa
Mie
M
Kuroiwa
Steven S S
SS
Poon
Tomio
T
Arai
Masashi
M
Toyoda
Hidenori
H
Akutsu
Akihiro
A
Umezawa
Ken-Ichi
K
Nakamura
Kaiyo
K
Takubo
D002881:Chromosomes, Human, Pair 12 / Q000235:genetics; D041321:Chromosomes, Human, X / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007621:Karyotyping; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000235:genetics*; D059505:Telomere Homeostasis / Q000235:genetics*; D014314:Trisomy / Q000235:genetics
D019098:Telomerase
Chromosome; Karyotype analysis; Q-FISH; Telomere; iPSC