Abstract
Human-induced pluripotent stem cells (hiPSCs) have shown great potential toward practical and scientific applications. We previously reported the generation of human dental pulp stem cells using non-integrating replication-defective Sendai virus (SeVdp) vector in feeder-free culture with serum-free medium hESF9. This study describes the generation of hiPSCs from peripheral blood mononuclear cells to increase the donor population, while reducing biopsy invasiveness. From 6-d-old primary culture of peripheral blood mononuclear cells (PBMCs) with IL-2, hiPSCs were established using SeVdp(KOSM)302L with recombinant Laminin-511 E8 fragments under serum-free condition. The established PBMC-derived hiPSCs showed pluripotency and differentiation ability both in vivo and in vitro. In addition, we evaluated microarray data from PBMC- and dental pulp–derived hiPSCs. These hiPSCs will be beneficial for characterizing the molecular mechanisms of cellular differentiation and may provide useful substrates for developing cellular therapeutics.
Highlights
To prevent the risk of contaminating Human-induced pluripotent stem cells (hiPSCs) with unknown viruses, unknown substances, and unpredictable genome insertions, as well as to standardize a culture method under defined conditions, we previously developed and reported culture systems capable of maintaining both the undifferentiated status and pluripotency of embryonic stem cells (ESCs) and iPSCs without using serum or feeder cells or retroviruses from dental pulp cells (DPCs) (Yamasaki et al 2016), eliminating the risk of activating nearby oncogenes by gene insertions
Reprogramming efficiencies The obtained peripheral blood mononuclear cells (PBMCs) on Days 0, 3, and 6 with interleukin-2 (IL-2) supplemented RD6F were used for hiPSC cell reprogramming with the SeVdp(KOSM)302L vector at an multiplicity of infection (MOI) of 6, using LamininE8 under completely feeder-free and serum-free culture conditions
We previously reported induction of hiPSCs from fibroblasts derived from gingiva or DPCs derived from extracted teeth (Yamasaki et al 2016; Yamasaki et al 2014)
Summary
Human-induced pluripotent stem cells (hiPSCs) (Takahashi and Yamanaka 2006) research holds tremendous potential for regenerative medicine, drug discovery, and disease modeling with the added advantage of circumventing the ethical concerns regarding the use of pluripotent cells derived from embryos. Murine-derived feeder cells are widely used to maintain the pluripotency of hiPSCs, and human-derived feeder cells are used. These cells are unsuitable for stem cell maintenance (Price 2017) as the feeder cells are cultured in medium supplemented with fetal bovine serum or proprietary serum replacements.
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