title: Optimized differentiation of hiPSC-derived Schwann cells and their integration into 2D and 3D neuromuscular tricultures
creator: Hörner, Sarah Janice
subject: 570
subject: 570 Life sciences
description: Neuromuscular junctions (NMJs) are synapses in the peripheral nervous system where signals are transmitted from spinal motoneurons to skeletal muscle fibers, enabling locomotion. Numerous in vitro models for NMJs have been developed to investigate their formation, function, and disease mechanisms. In addition to motoneurons and muscle fibers, the NMJ also consists of other cell types, in particular, glial cells of Schwann cell origin. Although these are critical for all of the aforementioned processes, Schwann cells have not yet been integrated into neuromuscular in vitro models in a targeted manner, primarily due to limitations regarding the efficient generation of human Schwann cells in vitro. To address this, an optimized protocol for robust and efficient generation of human induced pluripotent stem cell (hiPSC)-derived Schwann cells is presented in this thesis. To achieve this, a number of different modifications to a published differentiation protocol were tested. The activation of BMP signaling to a precisely tuned intermediate level proved to be important for the efficient generation of neural crest cells, which could be differentiated into Schwann cells subsequently. In addition, improved cell maturation was achieved using a defined culture medium, demonstrated by expression of typical Schwann cell marker proteins. This medium also promoted maturation of both hiPSC-derived motoneurons and murine cell line-derived myotubes, and was therefore compatible with neuromuscular tricultures. Using pre differentiated precursor cells, a protocol was established to combine all three cell types and culture them for several days. In such tricultures, Schwann cells aligned with motoneurons, and both were found to colocalize with nAChR clusters on myotubes and to influence their formation. Additionally, different approaches for multicellular and spatially organized neuromuscular cultures in 3D hydrogels were explored. Overall, this thesis provides the tools for selective integration of hiPSC-derived Schwann cells into neuromuscular cell culture models, which paves the way for future in vitro studies of NMJ formation and cell-type specific disease mechanisms.
date: 2023
type: Dissertation
type: info:eu-repo/semantics/doctoralThesis
type: NonPeerReviewed
format: application/pdf
identifier: https://archiv.ub.uni-heidelberg.de/volltextserverhttps://archiv.ub.uni-heidelberg.de/volltextserver/33522/1/Dissertation_SJHoerner.pdf
identifier: DOI:10.11588/heidok.00033522
identifier: urn:nbn:de:bsz:16-heidok-335228
identifier:   Hörner, Sarah Janice  (2023) Optimized differentiation of hiPSC-derived Schwann cells and their integration into 2D and 3D neuromuscular tricultures.  [Dissertation]     
relation: https://archiv.ub.uni-heidelberg.de/volltextserver/33522/
rights: info:eu-repo/semantics/openAccess
rights: http://archiv.ub.uni-heidelberg.de/volltextserver/help/license_urhg.html
language: eng