Abstract |
Introduction: Induced pluripotent stem cells (iPSC) that exhibit embryonic
stem cell-like properties with unlimited self-renewal and multilineage
differentiation properties, are a potential cell source in regenerative
medicine and cell-based therapy. Although retroviral and lentiviral
transduction methods to generate iPSC are well established, the risk of
mutagenesis limits the use of these products for therapeutic applications.
Materials and Methods: In this study, reprogramming of human dermal
fibroblasts (NHDF) into iPSC was carried out using non-integrative Sendai
virus for transduction. The iPSC clones were characterised based on the
morphological changes, gene expression of pluripotency markers, and
spontaneous and directed differentiation abilities into cells of different
germ layers. Results: On day 18-25 post-transduction, colonies with
embryonic stem cell-like morphology were obtained. The iPSC generated were
free of Sendai genome and transgene after passage 10, as confirmed by RT-
PCR. NHDF-derived iPSC expressed multiple pluripotency markers in qRT-PCR
and immunofluorescence staining. When cultured in suspension for 8 days,
iPSC successfully formed embryoid body-like spheres. NHDF-derived iPSC
also demonstrated the ability to undergo directed differentiation into
ectoderm and endoderm.
Conclusion: NHDF were successfully reprogrammed into iPSC using non-
integrating Sendai virus for transduction.
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