Mesenchymal Stromal Cells for Sphincter Regeneration: Role of Laminin Isoforms upon Myogenic Differentiation

Abstract

Multipotent mesenchymal stromal cells (MSCs) are well known for their tri-lineage potential and ability to differentiate in vitro into osteogenic, chondrogenic or adipogenic lineages. By selecting appropriate conditions MSCs can also be differentiated in vitro into the myogenic lineage and are therefore a promising option for cell-based regeneration of muscle tissue such as an aged or damaged sphincter muscle. For the differentiation into the myogenic lineage there is still a need to evaluate the effects of extracellular matrix proteins such as laminins (LM) which are crucial for different stem cell types and for normal muscle function. The laminin family consists of 16 functionally different isoforms with LM-211 being the most abundant isoform of adult muscle tissues. In the sphincter tissue a strong expression of the isoforms LM-211/221, LM-411/421 and LM-511/521 can be detected in the different cell layers. Bone marrow-derived MSCs in culture, however, mainly express the isoforms LM-411 and LM-511, but not LM-211. Even after myogenic differentiation, LM-211 can hardly be detected. All laminin isoforms tested (LM-211, LM-411, LM-511 and LM-521) showed a significant inhibition of the proliferation of undifferentiated MSCs but, with the exception of LM-521, they had no influence on the proliferation of MSCs cultivated in myogenic medium. The strongest cellular adhesion of MSCs was to LM-511 and LM-521, whereas LM-211 was only a weakly-adhesive substrate for MSCs. Myogenic differentiation of MSCs even reduced the interaction with LM-211, but it did not affect the interaction with LM-511 and LM-521. Since during normal myogenesis the latter two isoforms are the major laminins surrounding developing myogenic progenitors, α5 chain-containing laminins are recommended for further improvements of myogenic differentiation protocols of MSCs into smooth muscle cells.

Heterotypic Scaffold Design Orchestrates Primary Cell Organization and Phenotypes in Cocultured Small Diameter Vascular Grafts

“…Moreover, the initially seeded MSCs showed the desired arrangement, indicated by the elongated cytoskeletons and αSMA‐positive structures. Also collagen type IV was synthesized (Figure I), which was enclosing the individual vSMCs as a basement membrane‐like matrix (Figure S9, Supporting Information), mimicking the native situation . To prove that the high control over MEW fiber placement could also guide the orientation of the MSCs, they were monoseeded ( n = 4 MSC donors) on scaffolds i) without MEW layer, ii) an MEW layer with fiber winding angle of 30° (programmed angle 30°, measured angle 35.3° ± 0.8°), and iii) samples with MEW layer at an angle of 70° (programmed angle 70°, measured angle 71.6° ± 0.6°).…”

Precise injection of human mesenchymal stromal cells in the urethral sphincter complex of Göttingen minipigs without unspecific bulking effects

“…Recently we explored the composition of the extracellular matrix of the urethral tissue of Göttingen minipigs . Here we extended these studies and investigated the width of tissue layers and the presence of muscular tissue in the tissue layers in different zones of the porcine urethra (Fig.…”

Minocycline enhances the mesenchymal stromal/stem cell pro-healing phenotype in triple antimicrobial-loaded hydrogels

“…MSC attachment to fibrinogen has been shown to upregulate immunosuppression by inducing an increased secretion of IL-6 and indoleamine dioxygenase (IDO) while mitigating T cell proliferation [57]. MSC attachment to laminin controls proliferation and is observed in a myogenic differentiation study [58]. The synergistic effect of vancomycin and minocycline inducing increased attachment to the ECM proteins may increase MSC viability and wound healing functionality at the wound site that could potentially contribute to accelerated wound healing.…”