In vitro and in vivo arterial differentiation of human multipotent adult progenitor cells

Abstract

IntroductionThe vascular system is a bipolar complex network of arteries that transport oxygen-rich blood to all tissues and veins that bring oxygendeprived blood back to the heart. 1 Because of this bipolar set-up, arteries and veins feature anatomic and physiological differences. Unlike venous endothelium, arterial endothelium is surrounded by several layers of smooth muscle cells (SMCs), separated by elastic laminae, and embedded in a thick layer of fibrillar collagen. 2 Moreover, both vessel types have a differential susceptibility to atherosclerotic disease, possibly due to exposure to different levels of shear stress. Arterial and venous endothelial cells (ECs) also have a distinct molecular signature, and such molecular specification occurs before the onset of blood flow. 3 Indeed, arteriovenous (AV) specification of ECs is accomplished early in development and is associated with the expression of a specific complement of factors: venous endothelium is characterized by the expression of EphB4, 4 Lefty-1, 5 Lefty-2, 5 COUP-TFII, 6 and MYO1-␤, 5 and arterial ECs express high levels of Notch 1 and 4, 7 Dll-4, 8 EphrinB1 and EphrinB2, 4 Jagged-1 and -2, 7 connexin-40, and Hey-2 (gridlock zebrafish ortholog). 9,10 Studies in Xenopus, zebrafish, and mice have revealed that, besides blood flow, 11 vessel-intrinsic cues and-later in development-signals from outside the vasculature 12,13 are implicated in defining arterial or venous fate such as members of the TGF-␤ pathway, 14,15 VEGF isoforms,13,[16][17][18]17 angiopoietins, 19 the Notch pathway, 7,9,20-22 the patched pathway, 20 and COUP-TFII, a member of the orphan nuclear receptor superfamily. 6 Although it has been shown that some of these pathways are well conserved from zebrafish to mouse, less information is available on whether they have a similar role in humans. Because these molecular differences between arterial and venous ECs exist independently of blood flow and some of these factors work in an EC-intrinsic way, 2 it should be possible to manipulate some or all of these to endow ECs with an arterial or venous fate. Consistent with this notion, recent studies have suggested that arterial markers can be induced in primary mature ECs. 5,13,21,23,24 Many different stem cell populations, including bone marrow (BM) mononuclear cells, AC133 ϩ endothelial progenitor cells, and embryonic stem cells have the potential to differentiate in vitro and in vivo into mature and functional ECs. 4,[25][26][27][28] We have recently described another stem cell population, multipotent adult progenitor cells (MAPCs), that differentiates into most somatic cell types, including functional ECs, in vitro and in vivo. [29][30][31][32][33] The question of whether and how these stem cells can be coaxed into arterial or venous ECs has thus far not been addressed. In this study, we analyzed the in vitro and in vivo arterial and venous endothelial differentiation of human MAPCs (hMAPCs) and hAC133 ϩ cells. Materials and methodsAdditional and extended descriptions of methods are inc...

Multipotent adult progenitor cells sustain function of ischemic limbs in mice

“…GFP + mBMCs were isolated from the femurs, depleted of erythrocytes, washed, and transplanted. hMAPCs were established at the University of Navarra using BM from donors age 18-54, as described previously (46). Samples were obtained after informed consent was received from the donors according to guidelines of the Committee on the Use of Human Subjects in Research at the Clínica Universitaria, Pamplona.…”

“…For all stainings and analyses, 7-μm sections were used, spanning a muscle area of approximately 2 mm in thickness. H&E and Sirius red stainings were performed as described previously (46). For immunohistochemistry and immunofluorescence, a list of primary antibodies is provided as supplemental information (Supplemental Table 2).…”

Differentiation Potential of Human Postnatal Mesenchymal Stem Cells, Mesoangioblasts, and Multipotent Adult Progenitor Cells Reflected in Their Transcriptome and Partially Influenced by the Culture Conditions

“…Smooth muscle, skeletal muscle, and endothelial differentiation assays, as well as the in vitro and in vivo Matrigel assays were performed as previously described [14,23,27]. A detailed protocol of all differentiation and functional assays can be found in Supporting Information.…”

“…Even though hMAPCs are, like hMSCs and hMab, expanded adherent to culture vessels, differences in cell surface phenotype, expansion, and differentiation ability exist. For instance, hMAPCs can be expanded for more protracted periods of time than hMSCs and differentiate in vitro not only into mesenchymal cell types, but also into endothelial cells and cells with hepatocyte-like features [21][22][23][24]. Like hMSCs, hMAPCs have also significant trophic effects when grafted in vivo [25,26].…”

Cell-Based Vascularization Strategies for Skin Tissue Engineering

“…Other investigators reported the isolation of a different BM-derived multipotent cell type that displays a vast expansion capacity combined with a broad differentiation potential that encompasses cells from the three embryonic germ layers. 107 These multipotent adult progenitor cells (MAPC) are morphologically, molecularly, and functionally different from BM-MSC and can in vitro differentiate into EC that contribute to vasculogenesis in vivo [108][109][110][111] ( Table 1). Although MAPC represent an almost inexhaustible source of cells that can be used in STE vascularization, the longterm procedure required to derive MAPC makes an autologous approach unfeasible.…”