Controlled Growth Factor Delivery and Cyclic Stretch Induces a Smooth Muscle Cell-like Phenotype in Adipose-Derived Stem Cells

Brandan Walters,Paul A Turner,Bernd Rolauffs,Jan P Stegemann,Melanie L Hart

doi:10.3390/cells10113123

Brandan Walters, Paul A Turner + Show 3 more

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https://doi.org/10.3390/cells10113123

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Journal: Cells	Publication Date: Nov 11, 2021
Citations: 11	License type: CC BY 4.0

Affiliation: University of Michigan–Ann Arbor, University of Freiburg

Abstract

Adipose-derived stem cells (ASCs) are an abundant and easily accessible multipotent stem cell source with potential application in smooth muscle regeneration strategies. In 3D collagen hydrogels, we investigated whether sustained release of growth factors (GF) PDGF-AB and TGF-β1 from GF-loaded microspheres could induce a smooth muscle cell (SMC) phenotype in ASCs, and if the addition of uniaxial cyclic stretch could enhance the differentiation level. This study demonstrated that the combination of cyclic stretch and GF release over time from loaded microspheres potentiated the differentiation of ASCs, as quantified by protein expression of early to late SMC differentiation markers (SMA, TGLN and smooth muscle MHC). The delivery of GFs via microspheres produced large ASCs with a spindle-shaped, elongated SMC-like morphology. Cyclic strain produced the largest, longest, and most spindle-shaped cells regardless of the presence or absence of growth factors or the growth factor delivery method. Protein expression and cell morphology data confirmed that the sustained release of GFs from GF-loaded microspheres can be used to promote the differentiation of ASCs into SMCs and that the addition of uniaxial cyclic stretch significantly enhances the differentiation level, as quantified by intermediate and late SMC markers and a SMC-like elongated cell morphology.

Highlights

As the population ages, higher numbers of patients will need to be treated for diseases characterized by dysfunction of smooth muscle, including cardiovascular disease [1], and urinary incontinence [2]
Human adipose-derived mesenchymal stem cells isolated from adult normal, healthy subcutaneous adipose tissue (Rooster Bio, Frederick, MD, USA, Catalog #Mesenchymal stromal cells (MSCs)-021) were expanded in T175 flasks in Alpha Modification Essential Medium (Alpha MEM, Fisher, Rockville, MD, USA) supplemented with 10% qualified fetal bovine serum (FBS; Invitrogen, Carlsbad, CA, USA) and 1% penicillin and streptomyocin (PS; Invitrogen)
adipose-derived stem cells (ASCs) were seeded in 3D into collagen hydrogels along with (i) either no μspheres and no growth factors (Control), (ii) unloaded μspheres with growth factors in the media (GF), or (iii) with μspheres that had been loaded with growth factors (LS)

Summary

Introduction

Higher numbers of patients will need to be treated for diseases characterized by dysfunction of smooth muscle, including cardiovascular disease [1], and urinary incontinence [2]. In order to produce constructs to potentially treat affected tissues, a reliable and well-characterized source of smooth muscle cells (SMCs) is needed. Mesenchymal stromal cells (MSCs) offer a readily available source for these cells. Adipose-derived stem cells (ASCs) are increasingly being considered for their higher yield during extraction, ease of access, and decreased donor co-morbidity [3,4,5,6]. Adipose-derived progenitor cells already express some smooth muscle markers [7,8,9,10], suggesting that ASCs may naturally (in vivo) contribute to muscle development

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