title: Stretching human mesenchymal stromal cells on stiffness-customized collagen type I generates a smooth muscle marker profile without growth factor addition.
abstract: Using matrix elasticity and cyclic stretch have been investigated for inducing mesenchymal stromal cell (MSC) differentiation towards the smooth muscle cell (SMC) lineage but not in combination. We hypothesized that combining lineage-specific stiffness with cyclic stretch would result in a significantly increased expression of SMC markers, compared to non-stretched controls. First, we generated dense collagen type I sheets by mechanically compressing collagen hydrogels. Atomic force microscopy revealed a nanoscale stiffness range known to support myogenic differentiation. Further characterization revealed viscoelasticity and stable biomechanical properties under cyclic stretch with >99% viable adherent human MSC. MSCs on collagen sheets demonstrated a significantly increased mRNA but not protein expression of SMC markers, compared to on culture flasks. However, cyclic stretch of MSCs on collagen sheets significantly increased both mRNA and protein expression of α-smooth muscle actin, transgelin, and calponin versus plastic and non-stretched sheets. Thus, lineage-specific stiffness and cyclic stretch can be applied together for inducing MSC differentiation towards SMCs without the addition of recombinant growth factors or other soluble factors. This represents a novel stimulation method for modulating the phenotype of MSCs towards SMCs that could easily be incorporated into currently available methodologies to obtain a more targeted control of MSC phenotype.
authors: Siegfried Weller Institute for Trauma Research, BG Trauma Clinic Tuebingen, University of Tuebingen, Germany.
authors: Miriam
authors: M
authors: Rothdiener
authors: Department of Urology, University of Tuebingen, Germany.
authors: Miriam
authors: M
authors: Hegemann
authors: Siegfried Weller Institute for Trauma Research, BG Trauma Clinic Tuebingen, University of Tuebingen, Germany.
authors: Tatiana
authors: T
authors: Uynuk-Ool
authors: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA.
authors: Brandan
authors: B
authors: Walters
authors: Siegfried Weller Institute for Trauma Research, BG Trauma Clinic Tuebingen, University of Tuebingen, Germany.
authors: Piruntha
authors: P
authors: Papugy
authors: Siegfried Weller Institute for Trauma Research, BG Trauma Clinic Tuebingen, University of Tuebingen, Germany.
authors: Phong
authors: P
authors: Nguyen
authors: Siegfried Weller Institute for Trauma Research, BG Trauma Clinic Tuebingen, University of Tuebingen, Germany.
authors: Valentin
authors: V
authors: Claus
authors: Center for Medical Research, Medical University Clinic II, University of Tuebingen, Germany.
authors: Tanja
authors: T
authors: Seeger
authors: Clinic for Trauma and Restorative Surgery, BG Trauma Clinic Tuebingen, University of Tuebingen, Germany.
authors: Ulrich
authors: U
authors: Stoeckle
authors: Department of Orthopaedic Surgery, University of Tuebingen, Germany.
authors: Karen A
authors: KA
authors: Boehme
authors: Department of Urology, University of Tuebingen, Germany.
authors: Wilhelm K
authors: WK
authors: Aicher
authors: Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA.
authors: Jan P
authors: JP
authors: Stegemann
authors: Department of Orthopedics and Trauma Surgery, Albert-Ludwigs-University, Freiburg, Germany.
authors: Melanie L
authors: ML
authors: Hart
authors: Department of Anatomy, Christian-Albrechts-University, Kiel, Germany.
authors: Bodo
authors: B
authors: Kurz
authors: Center for Medical Research, Medical University Clinic II, University of Tuebingen, Germany.
authors: Gerd
authors: G
authors: Klein
authors: Department of Orthopedics and Trauma Surgery, Albert-Ludwigs-University, Freiburg, Germany.
authors: Bernd
authors: B
authors: Rolauffs
mesh_terms: D000199:Actins / Q000235:genetics; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D002135:Calcium-Binding Proteins / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D024042:Collagen Type I / Q000737:chemistry*; D005786:Gene Expression Regulation; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D008840:Microfilament Proteins / Q000235:genetics; D018625:Microscopy, Atomic Force; D009124:Muscle Proteins / Q000235:genetics; D009130:Muscle, Smooth / Q000166:cytology*; D010641:Phenotype; D014404:Tubulin / Q000378:metabolism; D000096985:Calponins
chemical_list: D000199:Actins; D015415:Biomarkers; D002135:Calcium-Binding Proteins; D024042:Collagen Type I; D008840:Microfilament Proteins; D009124:Muscle Proteins; D014404:Tubulin; D000096985:Calponins; C541116:ACTA2 protein, human; C051615:transgelin