Human amniotic fluid stem cell differentiation along smooth muscle lineage

Abstract

Functional smooth muscle engineering requires isolation and expansion of smooth muscle cells (SMCs), and this process is particularly challenging for visceral smooth muscle tissue where progenitor cells have not been clearly identified. Herein we showed for the first time that efficient SMCs can be obtained from human amniotic fluid stem cells (hAFSCs). Clonal lines were generated from c-kit(+) hAFSCs. Differentiation toward SM lineage (SMhAFSCs) was obtained using a medium conditioned by PDGF-BB and TGF-β1. Molecular assays revealed higher level of α smooth muscle actin (α-SMA), desmin, calponin, and smoothelin in SMhAFSCs when compared to hAFSCs. Ultrastructural analysis demonstrated that SMhAFSCs also presented in the cytoplasm increased intermediate filaments, dense bodies, and glycogen deposits like SMCs. SMhAFSC metabolism evaluated via mass spectrometry showed higher glucose oxidation and an enhanced response to mitogenic stimuli in comparison to hAFSCs. Patch clamp of transduced hAFSCs with lentiviral vectors encoding ZsGreen under the control of the α-SMA promoter was performed demonstrating that SMhAFSCs retained a smooth muscle cell-like electrophysiological fingerprint. Eventually SMhAFSCs contractility was evident both at single cell level and on a collagen gel. In conclusion, we showed here that hAFSCs under selective culture conditions are able to give rise to functional SMCs.

Serum‐free spheroid suspension culture maintains mesenchymal stem cell proliferation and differentiation potential

“…Finally, while this study was conducted with bone marrow derived MSCs, the same media formulations may have similar results with MSCs derived from different anatomic locations such as adipose, hair follicle, or amniotic fluid . In addition, recent studies developed high yield strategies to differentiate human iPSCs into MSCs that exhibited superior proliferation and differentiation potential than their adult counterparts .…”

“…Finally, while this study was conducted with bone marrow derived MSCs, the same media formulations may have similar results with MSCs derived from different anatomic locations such as adipose, hair follicle, or amniotic fluid. 20,21,[64][65][66][67] In addition, recent studies developed high yield strategies to differentiate human iPSCs into MSCs that exhibited superior proliferation and differentiation potential than their adult counterparts. 11 It will be interesting to explore whether the same media formulations can sustain the high proliferation and differentiation potential of iPSCderived MSCs, thereby providing an unlimited source of MSCs for cellular therapies.…”

Exogenous transforming growth factor-β1 enhances smooth muscle differentiation in embryonic mouse jejunal explants

“…Notably, however, intestines of mice lacking fibroblast growth factor 9 (FGF9) display premature myogenesis and upregulated TGFβ signalling (Geske et al, 2008). Furthermore, TGFβ1 has also been implicated in supporting SM differentiation in diverse other tissues including urinary bladder mesenchyme (Liu et al, 2010), amniotic stem cells (Ghionzoli et al, 2013) and neural crest cells (Huang et al, 2011).…”

Tissue-Engineered Vascular Grafts Created From Human Induced Pluripotent Stem Cells

“…Mesenchymal progenitors, as well as functional smooth muscle cells, have been derived from various diverse sources such as human hair follicle [11], amniotic fluid [12], bone marrow, adipose tissue [13], as well as human embryonic cells [14], and more recently from human induced pluripotent stem cells [15]. Several groups have used a method based on the spontaneous differentiation into embryoid bodies [16] from which smooth muscle cells are selected [17, 18].…”