Engineered Skeletal Muscle Bundles – Optimization of 3D Imaging Protocols
Lampela, Ella (2021)
Lampela, Ella
2021
Bioteknologian ja biolääketieteen tekniikan kandidaattiohjelma - Bachelor's Programme in Biotechnology and Biomedical Engineering
Lääketieteen ja terveysteknologian tiedekunta - Faculty of Medicine and Health Technology
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Hyväksymispäivämäärä
2021-05-07
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tuni-202104273919
https://urn.fi/URN:NBN:fi:tuni-202104273919
Tiivistelmä
Skeletal muscle is the most regenerative and abundant tissue of the human body, a heterogenous structure formed by aligned and multinucleated myotubes. It is a growing target of interest in the field of tissue engineering. Another topical interest in the field is using stem cells in engineered scaffold structures. Mesenchymal stem cells (MSCs) are indicated to have multilineage differentiation potential, as well as the ability to fuse with myogenic cells to create multinucleated myotubes (called hybrid myotubes). Studies suggest that coculturing MSCs with myogenic cells promotes the differentiation of MSCs towards myogenic lineage and their fusion into myotubes. Adipose-derived mesenchymal stem cells are indicated to potentially differentiate along the myogenic lineage. Engineering three-dimensional (3D) skeletal muscle structures utilizes a specially engineered mold, which creates a traction force lining the cells along the mechanical strain creating an oriented morphology which can be observed from the figures presented in the results of this study. When displaying results of three-dimensional experiment setups and samples, 3D image reconstruction and quantitative analysis of all three dimensions is necessary. Therefore, optimizing the staining protocol, imaging and presentation format for high-quality 3D reconstructed images is needed. The aim of this study is to improve the reliable detection of human adipose-derived stem cell (ASC) nuclei inside the myotubes when cocultured with mouse C2C12 cells. The reliable localization of nuclei inside the myotubes improves the reliability of fusion ratio determination. The 3D image resolution was enhanced through staining iterations and digital enhancing to a level, where quantitative 3D data analysis can begin, and the visual localization of nuclei inside the myotubes was possible.
Kokoelmat
- Kandidaatintutkielmat [8709]