Expansion and angiogenic potential of mesenchymal stem cells from patients with critical limb ischemia

• Type of Material: Article
• Authors: Luke Packard Brewster, Emory University; Scott Robinson, Emory University; Roy Wang, Emory University; Sarah Griffiths, Georgia Institute of Technology; Haiyan Li, Emory University; Alexandra Peister, Morehouse College; Ian Copland, Emory University; Todd McDevitt, Emory University
• Language: English
• Date: 2017-03-01
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2016
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jvs.2015.02.061
• Title of Journal or Parent Work: Journal of Vascular Surgery
• ISSN: 0741-5214
• Volume: 65
• Issue: 3
• Start Page: 826
• End Page: +
• Grant/Funding Information: NHLBI KO8HL119592 & Society for Vascular Surgery/American College of Surgeons Scientific Development Grant (LB),; American Heart Award Innovative Research Grant IRG14740001 (LB/IC),; Emory/Georgia Institute of Technology Regenerative Engineering and Medicine (LB, AP, IC, TM), which is supported in part by PHS Grant UL1TR000454 from the Clinical and Translational Science Award Program, National Institutes of Health, National Center for Advancing Translational Sciences.”; Emory Department of Surgery Startup Funds (LB).
• Keywords: HEART-DISEASE; THERAPY; PERIPHERAL ARTERIAL-DISEASE; MSCS; Surgery; NEOVASCULARIZATION CAPACITY; Peripheral Vascular Disease; DOUBLE-BLIND; MARROW MONONUCLEAR-CELLS; Cardiovascular System & Cardiology; CLINICAL-TRIALS; STROMAL CELLS; Science & Technology; Life Sciences & Biomedicine; PROGENITOR
• Research Categories: Health Sciences, Medicine and Surgery

Abstract

Background Critical limb ischemia (CLI) is a life- and limb-threatening condition affecting 1% to 10% of the population with peripheral arterial disease. Traditional revascularization options are not possible for up to 50% of CLI patients, in which case, the use of cellular therapies, such as bone marrow-derived mesenchymal stem cells (MSCs), hold great promise as an alternative revascularization therapy. However, no randomized, controlled phase 3 trials to date have demonstrated an improvement in limb salvage with cellular therapies. This may be due to poor cell quality (ie, inability to generate a sufficient number of angiogenic MSCs) or to the inadequate retention and viability of MSCs after delivery, or both. Because concerns remain about the expansion and angiogenic potential of autologous MSCs in the CLI population, the objective of this study was to examine the effect of our novel culture media supplement, pooled human platelet lysate (PL), in lieu of the standard fetal bovine serum (FBS), to improve the expansion potential of MSCs from CLI patients. We also characterized the in vitro angiogenic activity of MSCs from the tibia of amputated CLI limbs compared with MSCs from healthy donors. Methods MSCs were obtained from the tibia of four CLI patients (ISC) and four ISC patients with diabetes mellitus (ISC+DM) undergoing major amputation. Healthy MSCs were aspirated from the iliac crest of four young and healthy donors. MSCs were isolated and expanded in culture with PL or FBS. MSCs from passage 3 to 6 were used for phenotypic marker expression and for adipogenic and osteogenic differentiation and were tested for their in vitro angiogenic activity on human microdermal endothelial cells. In parallel MSCs were cultured to passage 11 for population-doubling calculations. Results MSCs from ISC and ISC+DM patients and from healthy patients exhibited appropriate expression of cell surface markers and differentiation capacity. Population doublings were significantly greater for PL-stimulated compared with FBS-stimulated MSCs in all groups. Biologically active amounts of angiogens were identified in the secretome of all MSCs without consistent trends among groups. PL expansion did not adversely affect the angiogenic activity of MSCs compared with FBS. The ISC and ISC+DM MSCs demonstrated angiogenic effects on endothelial cells similar to those of healthy and ISC MSCs. Conclusions PL promotes the rapid expansion of MSCs from CLI and healthy persons. Importantly, MSCs expanded from CLI patients demonstrate the desired angiogenic activity compared with their healthy counterparts. We conclude that autologous MSCs from CLI patients can be sufficiently expanded with PL and be expected to deliver requisite angiogenic effects in vivo. We expect the improved expansion of ISC and ISC+DM with PL to be helpful in improving the successful delivery of autologous MSCs to patients with CLI.