Assessment of the Therapeutic Potential of Endothelial Cells Differentiated from Human mRNA-transfected Induced Pluripotent Stem Cells in a Murine Model of Peripheral Arterial Disease

Abstract

Background: Autologous stem cell therapies have shown potential in promoting vascular regeneration. Induced pluripotent stem cells (iPSCs) are a novel stem cell type derived from somatic cell reprogramming and can be differentiated into endothelial cells (iPSC-ECs). Retroviral-transfected iPSC-ECs have been previously shown to promote angiogenesis, however such cells are not suitable for human studies. Therefore, we aimed here to assess the angiogenic potential of mRNA-transfected iPSC-ECs in a mouse model of peripheral arterial disease (PAD). Methods: iPSC-ECs were isolated by fluorescenceactivated cell sorting for endothelial marker CD31. To assess tube formation, iPSC-ECs were cultured on growth factor reduced Matrigel for 24 h. Male NOD-SCID mice were randomly assigned to receive iPSC-ECs or control saline. Unilateral femoral artery ligation was performed and cells were delivered via intramuscular injection at time of surgery. Recovery of the ischaemic limb was tracked using laser Doppler perfusion imaging on days 0, 1, 2, 4, 7, 10 and 14. At day 14, gastrocnemius and adductor muscle sampleswere taken for futurehistological analysis. Results: In vitro, iPSC-ECs promoted angiogenesis by forming tubular networks after 24 h of culture onMatrigel. In the hindlimb experiments, blood perfusion recovery in the ischaemic leg was significantly enhanced in animals that received iPSC-ECs, compared to controls (p< 0.05 at day 14).