Award Number : W 81 XWH - 11 - 1 - 0607 TITLE : iPSC - Derived MSCs that Are Genetically Engineered for Systemic Bone

The ability to efficiently generate integration-free induced pluripotent stem cells (iPSCs) from the most readily availablesource—peripheral blood—has the potential to expedite the advances of iPSC-based therapies. We have successfullygenerated integration-free iPSCs from cord blood (CB) CD34 cells with improved oriP/EBNA1-based episomal vectors (EV)using a strong spleen focus forming virus (SFFV) long terminal repeat (LTR) promoter. Here we show that Yamanaka factors(OCT4, SOX2, MYC, and KLF4)-expressing EV can also reprogram adult peripheral blood mononuclear cells (PBMNCs) intopluripotency, yet at a very low efficiency. We found that inclusion of BCL-XL increases the reprogramming efficiency byapproximately 10-fold. Furthermore, culture of CD3/CD19 cells or T/B cell-depleted MNCs for 4–6 days led to thegeneration of 20–30 iPSC colonies from 1 ml PB, an efficiency that is substantially higher than previously reported. PB iPSCsexpress pluripotency markers, form teratomas, and can be induced to differentiate in vitro into mesenchymal stem cells,cardiomyocytes, and hepatocytes. Used together, our optimized factor combination and reprogramming strategy lead toefficient generation of integration-free iPSCs from adult PB. This discovery has potential applications in iPSC banking,disease modeling and regenerative medicine. Citation: Su R-J, Baylink DJ, Neises A, Kiroyan JB, Meng X, et al. (2013) Efficient Generation of Integration-Free iPS Cells from Human Adult Peripheral Blood UsingBCL-XL Together with Yamanaka Factors. PLoS ONE 8(5): e64496. doi:10.1371/journal.pone.0064496 Editor: Zoran Ivanovic, French Blood Institute, France Received February 21, 2013; Accepted April 16, 2013; Published May 21, 2013 Copyright: 2013 Su et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by Loma Linda University (LLU) GRASP Award (XBZ), and Department of Defense (DOD) Concept Award W81XWH-11-1-0607(XBZ), United States Army Medical Research Acquisition Activity (USAMRAA) Grant W81XWH-12-1-0023 (DJB), and the Division of Anatomy, the Department ofBasic Sciences, the Center for Health Disparities and Molecular Medicine at LLU (KJP and RJS) and Radiation Research Laboratories in the Department of RadiationMedicine at LLU (DSG). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: [email protected].