Chromosomal Instability and Telomere Shortening in Long-Term Culture of Hematopoietic Stem Cells: Insights from a Cell Culture Model of RPS14 Haploinsufficiency

Abstract

The fate of cultivated primary hematopoietic stem cells (HSCs) with respect to genetic instability and telomere attrition has not yet been described in great detail. Thus, knowledge of the genetic constitution of HSCs is important when interpreting results of HSCs in culture. While establishing a cell culture model for myelodysplastic syndrome with a deletion in 5q by performing RPS14 knockdown, we found surprising data that may be of importance for any CD34+ cell culture experiments. We performed cytogenetic analyses and telomere length measurement on transduced CD34+ cells and untransduced control cells to observe the effects of long-term culturing. Initially, CD34+ cells had a normal median telomere length of about 12 kb and showed no signs of chromosomal instability. During follow-up, the median telomere length seemed to decrease and, simultaneously, increased chromosomal instability could be observed - in modified and control cells. One culture showed a clonal monosomy 7 - independent of prior RPS14 knockdown. During further culturing, it seemed that the telomeres re-elongated, and chromosomes stabilized, while TERT expression was not elevated. In summary, irrespective of our results of RPS14 knockdown in the long-term culture of CD34+ cells, it becomes clear that cell culture artefacts inducing telomere shortening and chromosomal instability have to be taken into account and regular cytogenetic analyses should always be performed.

Investigation of sperm telomere length as a potential marker of paternal genome integrity and semen quality

“…Please cite this article in press as: Federica Cariati, et al, Investigation of sperm telomere length as a potential marker of paternal genome integrity and semen quality, Reproductive BioMedicine Online (2016) Uncited references Thomay et al, 2014 7 Telomere length and DNA instability in human sperm Q13 502b s_bs_query 503b s_bs_query 504b s_bs_query 505b s_bs_query 506b s_bs_query 507b s_bs_query 508b s_bs_query 509b s_bs_query 510b s_bs_query 511b s_bs_query 512b s_bs_query 513b s_bs_query 514b s_bs_query 515b s_bs_query 516b s_bs_query 517b s_bs_query 518b s_bs_query 519b s_bs_query 520b s_bs_query 521b s_bs_query 522b s_bs_query 523b s_bs_query 524b s_bs_query 525b s_bs_query 526b s_bs_query 527b s_bs_query 528b s_bs_query 529b s_bs_query 530b s_bs_query 531b s_bs_query 532b s_bs_query 533b s_bs_query 534b s_bs_query 535b s_bs_query 536b s_bs_query 537b s_bs_query 538b s_bs_query 539b s_bs_query 540b s_bs_query 541b s_bs_query 542b s_bs_query 543b s_bs_query 544b s_bs_query 545b s_bs_query 546b s_bs_query 547b s_bs_query 548b s_bs_query 549b s_bs_query 550b s_bs_query 551b s_bs_query 552b s_bs_query 553b s_bs_query 554b s_bs_query 555b s_bs_query 556b s_bs_query 557b s_bs_query 558b s_bs_query 559b s_bs_query 560b s_bs_query 561b s_bs_query 562b s_bs_query 563b s_bs_query 564b s_bs_query 565b s_bs_query 8 F Cariati et al…”

Radiosensitivity in patients suffering from chronic kidney disease

“…Th e second point is that genomic instability has been estimated by using the net frequency of MN as a biomarker of genomic instability. Diff erent biomarkers have been proposed such as the use of the classical chromosomal aberration (Church et al 2012), telomere shortening (Moreno-Palomo et al 2014, Th omay et al 2014 or comet assay (Savina et al 2012); nevertheless, MN has a great biological relevance since it represents fi xed genetic damage resulting from both aneugenic and clastogenic mechanisms (Fenech 2007) and it is considered as a good surrogate marker of cancer risk (Bonassi et al 2011). Th e usefulness of the MN assay to evaluate individual ionizing radiation sensitivity has been reported to determine the intrinsic sensitivity to ionizing radiation in cancer patients, as well as in human immunodefi ciency virus (HIV)-positive individuals (Baeyens et al 2002, Mozdarani et al 2005, Christiakov et al 2008, Vral et al 2011, Bhatia and Kumar 2013.…”