Mouse embryonic stem cells that survive γ‐rays exposure maintain pluripotent differentiation potential and genome stability

Abstract

This study aimed to investigate the cell cycle, apoptosis, cytogenetics and differentiation capacity of mouse embryonic stem cells (mESCs) that survived a single dose of 2 or 5 Gy γ-rays during a period of up to 96 h of culture. After 2 Gy irradiation and 24 h culture, compared to control, a significant majority of cells was blocked at the G2/M phase and a massive apoptosis was recorded. Between 48 and 72 h post-irradiation, the parameters used to describe the cell cycle and apoptosis returned similar to those of control samples. When mESCs were irradiated with 5 Gy, a small fraction of cells, even after 96 h of culture, still presented clear evidences of a G2/M block and apoptosis. The cytogenetic analysis performed at 96 h showed that the structural stability of the aberrations did not change significantly when comparing control and 2 or 5 Gy-treated populations. However, the chromosomal damage observed in the progeny of the survived cells after 5 Gy exposure is significantly higher than that observed in control samples, although it is mostly of the stable and transmissible type. Ninety-six hours after irradiation, the survived mESCs maintained their undifferentiated status and capability to differentiate into the three germ layers. Overall, these results indicate a commitment of mESCs to maintain pluripotency and genome stability.

Arsenic trioxide alters the differentiation of mouse embryonic stem cell into cardiomyocytes

“…R1 mouse embryonic stem cell line (kindly provided by Dr. Nagy from Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada) and STO cell line were cultivated as previously described 55 .…”

“…Reverse transcription and quantitative Real-Time PCR reactions were performed as previously described 55 57 . The sequences of specific primers used were reported in Table 1S .…”

Heat shock induces apoptosis in human embryonic stem cells but a premature senescence phenotype in their differentiated progeny

“…32 Similar results were reported for mouse ESC survived γ-ray exposure. 33 hESC after irradiation in doses known to induce apoptosis of these cells are able to generate teratomas and express pluripotent genes. 34,35 In our experiments hESC exposed to sublethal heat shock easily underwent apoptosis, but survived cells rapidly restore their initial properties: morphology, high proliferation activity, pluripotent properties (expression of pluripotent markers, generation of embryoid bodies and differentiation into three germ layers).…”

Chromosomal Abnormalities in Embryonic and Somatic Stem Cells

“…This genomic instability does not affect their stemness and, upon challenging, SCs achieve a differentiated phenotype [Rebuzzini et al, 2012[Rebuzzini et al, , 2013Gaztelumendi and Nogués, 2014;Luft et al, 2014]. Strikingly, there is a lack of studies that analytically investigate if and which type of aneuploidies is permissive for cell differentiation, an aspect that is central to the use of SCs in the clinic.…”