Setting Up an Engineered Heart Tissue Model
Kulmala, Lotta (2024)
2024
Bioteknologian ja biolääketieteen tekniikan maisteriohjelma - Master's Programme in Biotechnology and Biomedical Engineering
Lääketieteen ja terveysteknologian tiedekunta - Faculty of Medicine and Health Technology
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Hyväksymispäivämäärä
2024-04-23
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tuni-202404033246
https://urn.fi/URN:NBN:fi:tuni-202404033246
Tiivistelmä
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) transformed cardiac research. Since then, efforts to create physiologically relevant models of the human heart have been constant. Traditional 2D hiPSC-CMs lack some important attributes of the myocardium, such as cell maturity, mechanobiological cues and 3D organisation. Tissue-engineered cardiac constructs have been developed to try to overcome these shortcomings. This thesis describes the process of integrating an Engineered Heart Tissue (EHT) model with previously used practices.
The EHT model set up in this thesis is a strip-shaped fibrin gel containing iPSC-CMs between silicone poles in a 24-well format. The iPSC-CMs are derived using a commonly used small molecule monolayer method, and the cardiomyocyte concentration was increased with metabolic lactate purification. The tissue constructs generate force under auxotonic stretch conditions, which is analysed video-optically. The morphology of the cells is visualised with immunocytochemical staining to detect signs of maturity.
Functioning EHTs could be produced in our laboratory, but the tissue strips did not reach a fully coherent beating. The lack of synchronisation is assumed to be due to too low cardiac differentiation efficiency. The immunostained samples revealed elongated and organised CMs in the tissue, expressing higher morphological maturity. The functional analysis showed improvements in the contraction kinetics as the hydrogel production was optimised, but the contractile forces remained at low levels. Despite some challenges, it is evident through this work that EHTs can be produced in our laboratory and are a promising platform for our current research topics.
The EHT model set up in this thesis is a strip-shaped fibrin gel containing iPSC-CMs between silicone poles in a 24-well format. The iPSC-CMs are derived using a commonly used small molecule monolayer method, and the cardiomyocyte concentration was increased with metabolic lactate purification. The tissue constructs generate force under auxotonic stretch conditions, which is analysed video-optically. The morphology of the cells is visualised with immunocytochemical staining to detect signs of maturity.
Functioning EHTs could be produced in our laboratory, but the tissue strips did not reach a fully coherent beating. The lack of synchronisation is assumed to be due to too low cardiac differentiation efficiency. The immunostained samples revealed elongated and organised CMs in the tissue, expressing higher morphological maturity. The functional analysis showed improvements in the contraction kinetics as the hydrogel production was optimised, but the contractile forces remained at low levels. Despite some challenges, it is evident through this work that EHTs can be produced in our laboratory and are a promising platform for our current research topics.