2018 Volume 64 Issue 3 Pages 207-215
Objective: Werner syndrome (WS) is a rare autosomal recessive disorder characterized by the premature onset of several aging-associated diseases. WS is caused by mutations in WRN, encoding a RecQ DNA helicase that plays a role in the maintenance of genomic stability. To elucidate the mechanisms underlying WS-associated aging, it is important to establish a WS experimental model. The objective of this study was to generate induced pluripotent stem cells (iPSCs) from cells derived from a patient with WS and to obtain differentiated cells from those iPSCs to study the mechanisms underlying WS-associated aging.
Methods: Peripheral blood was sampled from a patient with WS, and the T lymphocytes isolated from those samples were activated by IL-2 and anti-CD3 antibody. Next, the cells were transduced with reprogramming factors (OCT4, SOX2, KLF4, and MYC ) using Sendai virus, and WS-specific iPSCs (WS-iPSCs) were generated.
Results: The expression analysis and the teratoma formation assay revealed that the WS-iPSCs expressed pluripotency markers and differentiated into all tissues derived from all three germ layers. Importantly, WS-iPSCs showed normal karyotypes, with proliferation rates similar to that of control iPSCs. WS-iPSCs were culturable for over two years, maintaining their pluripotent status, and they differentiated into endothelial cells (ECs) and smooth muscle cells (SMCs) in vitro.
Conclusion: We generated WS-iPSCs with normal karyotypes, and these cells differentiated to ECs and SMCs, which could be studied to elucidate the mechanisms underlying premature aging in WS.
