Hokkaido University Graduate School of Medicine, Sapporo
Country
Japan
Year
2017
Journal
Dermatological Science
Abstract
Background: Induced pluripotent stem cell (iPSC) technology enables patient-specific pluripotent stem
cells to be derived from adult somatic cells without the use of an embryonic cell source. To date, recessive
dystrophic epidermolysis bullosa (RDEB)-specific iPSCs have been generated from patients using
integrating retroviral vectors. However, vector integration into the host genome can endanger the
biosafety and differentiation propensities of iPSCs. Although various integration-free reprogramming
systems have been reported, their utility in reprogramming somatic cells from patients remains largely
undetermined.
Objective: Our study aims to establish safe iPSCs from keratinocytes of RDEB patients using nonintegration
vector.
Method: We optimized and infected non-integrating Sendai viral vectors to reprogram keratinocytes from
healthy volunteers and RDEB patients.
Results: Sendai vector infection led to the reproducible generation of genomic modification-free iPSCs
from these keratinocytes, which was proved by immunohistochemistry, reverse transcription
polymerase chain reaction, methylation assay, teratoma assay and embryoid body formation assay.
Furthermore, we confirmed that these iPSCs have the potential to differentiate into dermal
fibroblasts
and epidermal keratinocytes.
Conclusion: This is the
first report to prove that the Sendai vector system facilitates the reliable
reprogramming of patient keratinocytes into transgene-free iPSCs, providing another pluripotent
platform for personalized diagnostic and therapeutic approaches to RDEB.