Attenuation of Hind-Limb Ischemia in Mice with Endothelial-Like Cells Derived from Different Sources of Human Stem Cells

Abstract

Functional endothelial-like cells (EC) have been successfully derived from different cell sources and potentially used for treatment of cardiovascular diseases; however, their relative therapeutic efficacy remains unclear. We differentiated functional EC from human bone marrow mononuclear cells (BM-EC), human embryonic stem cells (hESC-EC) and human induced pluripotent stem cells (hiPSC-EC), and compared their in-vitro tube formation, migration and cytokine expression profiles, and in-vivo capacity to attenuate hind-limb ischemia in mice. Successful differentiation of BM-EC was only achieved in 1/6 patient with severe coronary artery disease. Nevertheless, BM-EC, hESC-EC and hiPSC-EC exhibited typical cobblestone morphology, had the ability of uptaking DiI-labeled acetylated low-density-lipoprotein, and binding of Ulex europaeus lectin. In-vitro functional assay demonstrated that hiPSC-EC and hESC-EC had similar capacity for tube formation and migration as human umbilical cord endothelial cells (HUVEC) and BM-EC (P>0.05). While increased expression of major angiogenic factors including epidermal growth factor, hepatocyte growth factor, vascular endothelial growth factor, placental growth factor and stromal derived factor-1 were observed in all EC cultures during hypoxia compared with normoxia (P<0.05), the magnitudes of cytokine up-regulation upon hypoxic were more dramatic in hiPSC-EC and hESC-EC (P<0.05). Compared with medium, transplanting BM-EC (n = 6), HUVEC (n = 6), hESC-EC (n = 8) or hiPSC-EC (n = 8) significantly attenuated severe hind-limb ischemia in mice via enhancement of neovascularization. In conclusion, functional EC can be generated from hECS and hiPSC with similar therapeutic efficacy for attenuation of severe hind-limb ischemia. Differentiation of functional BM-EC was more difficult to achieve in patients with cardiovascular diseases, and hESC-EC or iPSC-EC are readily available as “off-the-shelf” format for the treatment of tissue ischemia.

Specific Role of Impaired Glucose Metabolism and Diabetes Mellitus in Endothelial Progenitor Cell Characteristics and Function

“…In contrast to early EPCs, late EPC cultures are more difficult to establish and sometimes impossible in some patients with DM and cardiovascular disease. 13 Many clinical studies have demonstrated that number of circulating EPCs is reduced and their function impaired in terms of decreased proliferation, adhesion, and vasculogenesis in patients with type 1 and type 2 DM (Table I in the online-only Data Supplement). Similar alterations in EPCs are observed in patients with type 1 and type 2 DM, suggesting a close mechanistic link between hyperglycemia and the reduction and functional impairment of EPCs.…”

“…An exogenous source of EPCs such as those derived from pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, can avoid this issue. 13 Nonetheless, other issues related to the use of such cell sources, such as risk of immune rejection and tumor formation, need to be addressed.…”

MicroRNA let‐7g possesses a therapeutic potential for peripheral artery disease

“…Mice then were divided into two groups (n = 6 for each group) for intramuscular injection (IM) of 5 nM negative control-miRNA (placebo group) or let-7g mimic (let-7g group). IM injection was given at three sites of the gastrocnemius muscle at the medial thigh on day 1, 8 and 15 after induced hindlimb ischaemia [14]. Noticeably, the day for induction of ischaemic limb was defined as day 0.…”

Generating induced pluripotent stem cell derived endothelial cells and induced endothelial cells for cardiovascular disease modelling and therapeutic angiogenesis

“…Preliminary cell survival results showed iPSC-EC numbers were greatly reduced by day 7 and undetectable by day 11, which prompted the inclusion of an additional cell treatment at day 7 [6]. In a study comparing the therapeutic efficacy of human iPSC-ECs relative to bone marrow derived ECs, ESC-ECs and human umbilical cord ECs (HUVECs), Lai et al found that treatment with iPSC-ECs enhanced reperfusion in the ischaemic limb as effectively as ESC-ECs and bone marrow derived ECs and more effectively than HUVECs [93].…”