Pharmacological priming of adipose-derived stem cells for paracrine VEGF production with deferoxamine Guei-Sheung Liu 1,2 *, Hitesh M. Peshavariya 1,2 , Masayoshi Higuchi 1 , Elsa C. Chan 1,2 , Gregory J. Dusting 1,2 and Fan Jiang 1,3 * 1 O’Brien Institute, Fitzroy, Victoria, Australia 2 Centre for Eye Research Australia and Department of Ophthalmology, University of Melbourne, East Melbourne, Victoria, Australia 3 Key Laboratory of Cardiovascular Remodeling and Function Research, Qilu Hospital, Shandong University, Jinan, Shandong Province, China Abstract Adipose-derived stem cells (ASCs) show great potentials in applications such as therapeutic angio- genesis, regenerative medicine and tissue engineering. Pharmacological preconditioning of stem cells to boost the release of cytoprotective factors may represent an effective way to enhance their therapeutic efficacy. In this study, the aim was to determine whether deferoxamine can enhance the release of vascular endothelial growth factor (VEGF) from in vitro expanded ASCs. It is demon- strated that deferoxamine (50–300 μM) upregulated VEGF expression in a concentration- and time-dependent fashion. At the concentrations used, deferoxamine did not show any cytotoxic effects. The stimulatory effect of deferoxamine on VEGF expression was mediated by augmentation of hypoxia inducible factor-1 in ASCs, but independent of its antioxidant properties. Moreover, deferoxamine enhanced the paracrine effects of ASCs in promoting the regenerative functions of endothelial cells (migration and in vitro wound healing activities). This study provides evidence that deferoxamine might be a useful drug with low cell toxicity for pharmacological preconditioning of ASCs to enhance their capacity of VEGF production. Copyright © 2013 John Wiley & Sons, Ltd. Received 25 July 2012; Revised 8 February 2013; Accepted 12 June 2013 Supporting information may be found in the online version of this article. Keywords adipose-derived stem cells; deferoxamine; hypoxia inducible factor-1; paracrine; pharmacolog- ical preconditioning; vascular endothelial growth factor 1. Introduction Adipose-derived stem cells (ASCs) were originally identified from the stromal vascular fraction of aspirated fat tissue during liposuction surgery (Zuk et al., 2002). These cells have been shown to possess multi-lineage differentiation potential to generate fat, bone, cartilage, vascular and muscle tissues following directed differentiation (Gimble et al., 2007; Locke et al., 2009). Because of the large amount of ASCs available in adult tissues and the ease of obtaining them in comparison with other stem cell sources such as bone marrow, a great deal of attention has been given to their application in tissue engineering and regenerative medicine (Kuhbier et al., 2010). For example, a number of animal and early clinical studies have demonstrated that ASC transplantation may be an effective therapy for ameliorating cardiac dysfunction and preventing adverse cardiac remodelling after myocardial infarction (Sanz-Ruiz et al., 2009; Hong et al., 2010). Accumulating evidence has suggested that ASCs may exert beneficial effects by producing a number of cytoprotective factors (cytokines and growth factors) *Correspondence to: F. Jiang, Key Laboratory of Cardiovascular Remodeling and Function Research, Qilu Hospital, 107 Wen Hua Xi Road, Jinan 250012, China. E-mail: fjiang@sdu.edu.cn G.-S. Liu, Centre for Eye Research Australia & Department of Oph- thalmology, University of Melbourne Level 1, 32 Gisborne Street, East Melbourne, VIC 3002, Australia. E-mail: guei-sheung. liu@unimelb.edu.au Copyright © 2013 John Wiley & Sons, Ltd. JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE RESEARCH ARTICLE J Tissue Eng Regen Med (2013) Published online in Wiley Online Library (wileyonlinelibrary.com) DOI: 10.1002/term.1796