Abstract
In this study we demonstrate that CD34+ cells derived from human embryonic stem cells (hESCs) have higher smooth muscle cell (SMC) potential than CD34− cells. We report that from all inductive signals tested, retinoic acid (RA) and platelet derived growth factor (PDGFBB) are the most effective agents in guiding the differentiation of CD34+ cells into smooth muscle progenitor cells (SMPCs) characterized by the expression of SMC genes and proteins, secretion of SMC-related cytokines, contraction in response to depolarization agents and vasoactive peptides and expression of SMC-related genes in a 3D environment. These cells are also characterized by a low organization of the contractile proteins and the contractility response is mediated by Ca2+, which involves the activation of Rho A/Rho kinase- and Ca2+/calmodulin (CaM)/myosin light chain kinase (MLCK)-dependent pathways. We further show that SMPCs obtained from the differentiation of CD34+ cells with RA, but not with PDGFBB, can be maturated in medium supplemented with endothelin-1 showing at the end individualized contractile filaments. Overall the hESC-derived SMCs presented in this work might be an unlimited source of SMCs for tissue engineering and regenerative medicine.
Highlights
Vascular smooth muscle cells (VSMCs) have enormous applications in regenerative medicine [1,2,3]
We show that CD34+ cells have higher SMC potential than CD342 cells and PDGFBB and retinoic acid (RA) are the most effective agents to drive the differentiation of human embryonic stem cells (hESCs) into smooth muscle progenitor cells (SMPCs)
We further show that RA or PDGFBB drive the differentiation of CD34+ cells into SMPCs We have characterized the differentiated cells at gene and at protein level, their secretome, the ability to contract when incubated with several pharmacological agents, and contraction mechanism mediated by Ca2+
Summary
Vascular smooth muscle cells (VSMCs) have enormous applications in regenerative medicine [1,2,3]. SMLCs transplanted subcutaneously in an animal model were able to contribute for the formation of functional blood microvessels [12]. Despite these advances, several issues remain poorly understood: (i) what hESC-derived population has the most SMC potential, (ii) the bioactive molecules involved in the differentiation process, (iii) the modulatory effect of 3D environments in SMLCs, (iv) the functionality of the differentiated SMLCs, and (v) the level of organization of the contractile protein filaments
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