| dc.contributor.advisor | Zimmermann, Wolfram-Hubertus Prof. Dr. | |
| dc.contributor.author | Shahriyari, Mina | |
| dc.date.accessioned | 2021-06-25T13:47:31Z | |
| dc.date.available | 2021-07-02T00:50:07Z | |
| dc.date.issued | 2021-06-25 | |
| dc.identifier.uri | http://hdl.handle.net/21.11130/00-1735-0000-0008-587F-8 | |
| dc.identifier.uri | http://dx.doi.org/10.53846/goediss-8688 | |
| dc.language.iso | eng | de |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject.ddc | 610 | |
| dc.title | Engineered skeletal muscle from human pluripotent stem cells to model muscle disease and regeneration | de |
| dc.type | doctoralThesis | de |
| dc.contributor.referee | Linke, Wolfgang Prof. Dr. | |
| dc.date.examination | 2021-02-09 | |
| dc.description.abstracteng | Skeletal muscle disease modeling offers the unique opportunity to investigate devastating muscle diseases like Duchenne Muscle Dystrophy in vitro but requires advanced three-dimensional (3D) model systems reflecting the characteristics of human muscle in vivo. The aim of this study was to generate engineered models of skeletal muscle from human pluripotent stem cells (hPSCs) with physiological properties by recapitulating specific stages of muscle development.
To allow for robust skeletal muscle tissue engineering first a directed differentiation protocol was established in 2D culture under serum-free conditions. Comparison of hPSC differentiation to embryonic muscle development confirmed significant overlap with characteristic signatures of paraxial mesoderm, dermatomyotome, and somite stage. The protocol robustly directed multiple hPSC lines into skeletal muscle cells in 2D culture as well as in a collagen-1/Matrigel® hydrogel in 3D generating bioengineered skeletal muscle (BSM) organoid. By identifying additional maturation cues (creatine, triiod-L-thyronine) hPSC-derived skeletal myogenic cells embedded into a collagen-1/Matrigel® hydrogel generated engineered skeletal muscle (ESM) with compact muscle syncytia, anisotropically arranged sarcomeres, properly localized dystrophin-associated complex proteins, and contractile function of developing fast muscle. Importantly, Pax7-positive cells were found adjacent to muscle fibers underneath a laminin-positive basal lamina in a satellite-like cell position. Cardiotoxin injury of ESM induced a regenerative response with recovery of tetanic force after complete loss of function. Finally, modelling of Duchenne Muscular Dystrophy (DMD) in ESM demonstrates “proof of concept” for efficacy of CRISPR/Cas9 based exon skipping.
Collectively, human BSM and ESM models provide unprecedented opportunities to study muscle development, maturation, and regeneration in vitro and may serve as preclinical test bed for novel therapies of skeletal muscle disease. | de |
| dc.contributor.coReferee | Fischer, André Prof. Dr. | |
| dc.subject.eng | Skeletal muscle organoid, tissue engineering, limb muscle, hypaxial dermomyotome, satellite cells, regeneration | de |
| dc.identifier.urn | urn:nbn:de:gbv:7-21.11130/00-1735-0000-0008-587F-8-4 | |
| dc.affiliation.institute | Medizinische Fakultät | |
| dc.subject.gokfull | Medizin (PPN619874732) | de |
| dc.subject.gokfull | Pharmakologie / Toxikologie / Pharmakotherapie - Allgemein- und Gesamtdarstellungen (PPN61987550X) | de |
| dc.description.embargoed | 2021-07-02 | |
| dc.identifier.ppn | 1761310348 | |