Smooth Muscle-Like Cells Generated from Human Mesenchymal Stromal Cells Display Marker Gene Expression and Electrophysiological Competence Comparable to Bladder Smooth Muscle Cells

Brun, Juliane; Lutz, Katrin A.; Neumayer, Katharina M. H.; Klein, Gerd; Seeger, Tanja; Uynuk-Ool, Tatiana; Wörgötter, Katharina; Schmid, Sandra L.; Kraushaar, Udo; Guenther, Elke; Rolauffs, Bernd; Aicher, Wilhelm K.; Hart, Melanie L.

doi:10.1371/journal.pone.0145153

Cited by 31 publications

(28 citation statements)

References 67 publications

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“…αSMA and VMT are known as common differentiation markers of smooth muscle cells, while MYH11 and CNN1 are late differentiation markers and more specific to contractile smooth muscle cells. [ 37 ] Expression of MYH11 and CNN1 in the tissues over time (Figure 6C,D) revealed that hiSMCs were differentiated into contractile phenotype in the aligned scaffolds and cytologically capable of performing potential contraction/relaxation. We also detected the expression of type I collagen (COL1A1), as the differentiation of SMCs induces the deposition of ECM proteins including collagen.…”

Section: Resultsmentioning

confidence: 99%

Bi‐Layered Tubular Microfiber Scaffolds as Functional Templates for Engineering Human Intestinal Smooth Muscle Tissue

Chen

Guo

Manousiouthakis

et al. 2020

Adv Funct Materials

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Designing biomimetic scaffolds with in vivo-like microenvironments using biomaterials is an essential component of successful tissue engineering approaches. The intestinal smooth muscle layers exhibit a complex tubular structure consisting of two concentric muscle layers in which the inner circular layer is orthogonally oriented to the outer longitudinal layer. Here, a 3D bi-layered tubular scaffold is presented based on flexible, mechanically robust, and well aligned silk protein microfibers to mimic the native human intestinal smooth muscle structure. The scaffolds are seeded with primary human intestinal smooth muscle cells to replicate intestinal muscle tissues in vitro. Characterization of the tissue constructs reveals good biocompatibility and support for cell alignment and elongation in the different scaffold layers to enhance cell differentiation and functions. Furthermore, the engineered smooth muscle constructs support oriented neurite outgrowth, a requisite step to achieve functional innervation. These results suggest these microfiber scaffolds as functional templates for in vitro regeneration of human intestinal smooth muscle systems. The scaffolding provides a crucial step toward engineering functional human intestinal tissue in vitro, as well as engineering other types of smooth muscles in terms of their similar phenotypes. Such utility may lead to a better understanding of smooth muscle associated diseases and treatments.

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Section: Resultsmentioning

confidence: 99%

Bi‐Layered Tubular Microfiber Scaffolds as Functional Templates for Engineering Human Intestinal Smooth Muscle Tissue

Chen

Guo

Manousiouthakis

et al. 2020

Adv Funct Materials

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“…SMCs in adult blood vessels exhibit low rate of proliferation, low synthetic activity, and expression of distinctive SMC markers such as a-SM-actin, SM-myosin, and other SMspecific markers, including calponin. 44,45 SMCs in vascular grafts exposed to stiffer matrices tend to undergo dedifferentiation, and transit into a pro-proliferative myofibroblastlike phenotype, particularly at the site of suture. 46,47 The difference in the elasticity of the native artery and the vascular graft at the site of suturing is called compliance mismatch.…”

Section: Discussionmentioning

confidence: 99%

Development of an electrospun biomimetic polyurea scaffold suitable for vascular grafting

et al. 2017

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The optimization of biomechanical and biochemical properties of a vascular graft to render properties relevant to physiological environments is a major challenge today. These critical properties of a vascular graft not only regulate its stability and integrity, but also control invasion of cells for scaffold remodeling permitting its integration with native tissue. In this work, we have synthesized a biomimetic scaffold by electrospinning a blend of a polyurea, poly(serinol hexamethylene urea) (PSHU), and, a polyester, poly-ε-caprolactone (PCL). Mechanical properties of the scaffold were varied by varying polymer blending ratio and electrospinning flow rate. Mechanical characterization revealed that scaffolds with lower PSHU content relative to PCL content resulted in elasticity close to native mammalian arteries. We also found that increasing electrospinning flow rates also increased the elasticity of the matrix. Optimization of elasticity generated scaffolds that enabled vascular smooth muscle cells (SMCs) to adhere, grow and maintain a SMC phenotype. The 30/70 scaffold also underwent slower degradation than scaffolds with higher PSHU content, thereby, providing the best option for in vivo remodeling. Further, Gly-Arg-Gly-Asp-Ser (RGD) covalently conjugated to the polyurea backbone in 30/70 scaffold resulted in significantly increased clotting times. Reducing surface thrombogenicity by the conjugation of RGD is critical to avoiding intimal hyperplasia. Hence, biomechanical and biochemical properties of a vascular graft can be balanced by optimizing synthesis parameters and constituent components. For these reasons, the optimized RGD-conjugated 30/70 scaffold electrospun at 2.5 or 5 mL/h has great potential as a suitable material for vascular grafting applications.

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“…We previously showed that P/PL is capable of inducing a smooth muscle-like phenotype in human bmMSCs [14,15]. The goal of the present study was to measure whether human P/PL alone was also sufficient in inducing a skeletal myogenic and neurogenic phenotype.…”

Section: Introductionmentioning

confidence: 90%

“…However, the overall efficacy and clinical endpoints were largely inconsistent. Such inconsistencies could be attributed to the dosage of injected cells, number of injections, delivery route, cell survival in situ after injection, endpoints examined, or timing of followup, but could also simply be the result of how the MSCs were prepared or expanded prior to application, as we have shown that expansion conditions can impact MSC lineage determination and prime the cells towards certain phenotypes [14,15].…”

Section: Introductionmentioning

confidence: 99%

Lack of a skeletal muscle phenotype in adult human bone marrow stromal cells following xenogeneic-free expansion

et al. 2020

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Background: Many studies have elegantly shown that murine and rat bone marrow-derived mesenchymal stromal cells (bmMSCs) contribute to muscle regeneration and improve muscle function. Yet, the ability of transplanted human bmMSCs to manifest myogenic potential shows conflicting results. While human adipose-and umbilical cord-derived MSCs can be differentiated into a skeletal muscle phenotype using horse serum (HS), bmMSCs have only been shown to differentiate towards the skeletal muscle lineage using a complex mixture of cytokines followed by transfection with notch intracellular domain. Methods: Since xenogeneic-free growth supplements are increasingly being used in the expansion of bmMSCs in clinical trials, we investigated the effects of human plasma and platelet lysate (P/PL) on the expression of neuromuscular markers and whether P/PL-expanded human bmMSCs could be differentiated towards a skeletal myogenic phenotype. Neuromuscular markers were measured using the highly sensitive droplet digital polymerase chain reaction for measuring the expression of Myf5, MyoD, MyoG, ACTA1, Desmin, GAP-43, and Coronin 1b transcripts, by performing immunofluorescence for the expression of Desmin, GAP-43, and MEF2, and flow cytometry for the expression of CD56/neural cell adhesion molecule (NCAM). Results: Despite that bmMSCs expressed the myogenic regulatory factor (MRF) MEF2 after expansion in P/PL, bmMSCs cultured under such conditions did not express other essential MRFs including Myf5, MyoD, MyoG, or ACTA1 needed for myogenesis. Moreover, HS did not induce myogenesis of bmMSCs and hence did not induce the expression of any of these myogenic markers. P/PL, however, did lead to a significant increase in neurogenic GAP-43, as well as Desmin expression, and resulted in a high baseline expression of the neurogenic gene Coronin 1b which was sustained under further P/PL or HS culture conditions. Fetal bovine serum resulted in equally high levels of GAP-43 and Coronin 1b. Moreover, the proportion of CD56/NCAM-positive bmMSCs cultured in P/PL was 5.9 ± 2.1. Conclusions: These data suggest that P/PL may prime a small portion of bmMSCs towards an early neural precursor cell type. Collectively, this shows that P/PL partially primes the cells towards a neurogenic phenotype, but does not prime adult human bmMSCs towards the skeletal muscle lineage.

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Smooth Muscle-Like Cells Generated from Human Mesenchymal Stromal Cells Display Marker Gene Expression and Electrophysiological Competence Comparable to Bladder Smooth Muscle Cells

Cited by 31 publications

References 67 publications

Bi‐Layered Tubular Microfiber Scaffolds as Functional Templates for Engineering Human Intestinal Smooth Muscle Tissue

Bi‐Layered Tubular Microfiber Scaffolds as Functional Templates for Engineering Human Intestinal Smooth Muscle Tissue

Development of an electrospun biomimetic polyurea scaffold suitable for vascular grafting

Lack of a skeletal muscle phenotype in adult human bone marrow stromal cells following xenogeneic-free expansion

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