Directed Differentiation of Skin-Derived Precursors Into Functional Vascular Smooth Muscle Cells

Abstract

Objective-The goal of this study was to characterize the factors and conditions required for smooth muscle cell (SMC)-directed differentiation of Sox2 ϩ multipotent rat and human skin-derived precursors (SKPs) and to define whether they represent a source of fully functional vascular SMCs for applications in vivo. Methods and Results-We found that rat SKPs can differentiate almost exclusively into SMCs by reducing serum concentrations to 0.5% to 2% and plating them at low density. Human SKPs derived from foreskin required the addition of transforming growth factor-␤1 or -␤3 to differentiate into SMCs, but they did so even in the absence of serum. SMC formation was confirmed by quantitative reverse transcription-polymerase chain reaction, immunocytochemistry, and fluorescence-activated cell sorting, with increased expression of smoothelin-B and little to no expression of telokin or smooth muscle ␥-actin, together indicating that SKPs differentiated into vascular rather than visceral SMCs. Rat and human SKP-derived SMCs were able to contract in vitro and also wrap around and support new capillary and larger blood vessel formation in angiogenesis assays in vivo. Conclusion-SKPs are Sox2ϩ progenitors that represent an attainable autologous source of stem cells that can be easily differentiated into functional vascular SMCs in defined serum-free conditions without reprogramming. SKPs represent a clinically viable cell source for potential therapeutic applications in neovascularization. (Arterioscler Thromb Vasc Biol. 2011;31:2938-2948.)

Skin-Derived Precursor Cells Promote Wound Healing in Diabetic Mice

“…A previous study 28 reported that SKPs directly differentiate vascular smooth muscle cells with the addition of transforming growth factor A in vivo. However, in our histological analysis of immunofluorescent stained region by PKH26 and antiYCD31 antibody, direct differentiation to vessels of applied SKPs was not confirmed (data not shown).…”

Regulation of skin aging and heart development by TAp63

“…Cardiac neural crest contributes to the heart stem cell pool 68 and the embryonic branchial arches (where TAp63 is detected) contain mesendodermal cells of neural crest origin that participate to the heart development. 69 As SKPs cells are also derived from the neural crest and can produce functional smooth muscle cells, 70 it would be interesting to determine whether the TAp63-expressing endodermal cells with cardiogenic inductive role 36 have a common embryonic origin with the dermal SKPs and whether the latter could have cardiogenic activity on mesocardiac progenitors. Again, a careful analysis of the microarray and Chip-seq analysis of SKPs cells may be informative.…”

Differentiation and Application of Induced Pluripotent Stem Cell–Derived Vascular Smooth Muscle Cells

“…98 Thus, alternative cell types, such as skin-derived precursors, ought to be recognized for their potential as VSMC sources as they readily differentiate into VSMCs using the same growth factors required to generate VSMCs from neural crest precursors. 99 Moreover, they have been used to model type II diabetes mellitus in elderly patients, without the need for reprogramming required in iPSC generation, often resulting in erasure of disease-relevant epigenetic modifications. 100 Drug screening may also prove difficult using newly differentiated cells if pharmacotherapy is only required in the later stages of disease because newly differentiated cells will not have been exposed to environmental factors responsible for advancing pathological processes.…”