Cardiac extracellular matrix gel for ESC-derived cardiomyocyte maturation
Date
2020-08-31
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Abstract
The need for precision medicine has been established and the use of organoids to test individual drug efficacy and develop individual disease models is creating an opportunity for innovation. Human embryonic stem cells (hESCs) are commonly used for these applications but differentiation often results in immature phenotypes that may not be physiologically relevant. While hESCs can be differentiated into any cell type using small molecules, emerging concepts suggest that additional exogenous factors influence differentiation. In situ, stem cells exist in an organ-specific niche which contains extracellular matrix (ECM) proteins that have important paracrine effects to promote the maturity. This project aimed to investigate whether either of two alternative preparations of organ-specific ECM matrices support would offer improved support for tissue-specific differentiation and maturation. ECM matrices from porcine hearts (i.e. cardiogel) were prepared and decellularized either completely or partially. These cardiogel preparations were then studied using mass spectrometry and cytochemical staining before being used to support the maturation of hESCs that had been differentiated into cardiomyocytes (CMs). Moderate decellularization of porcine heart was achieved as shown by histochemical staining and the cardiogels contained variable amounts of important ECM components. However, neither cardiogel preparations appeared to increase the maturity of hESC-derived CMs, as assessed by cellular gene expression after 4 days of culture on cardiogel following differentiation. In conclusion, a protocol to generate cardiogels from porcine heart was achieved but significant batch variability was found. Further optimization may lead to a substrate which promotes cellular maturation and which can be used in conjunction with other technologies to increase cellular yield and homogeneity of resulting cardiac lineages.
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Keywords
Stem cell, Decellularization, Maturation, Differentiation
Citation
Thompson, M. J. (2020). Cardiac extracellular matrix gel for ESC-derived cardiomyocyte maturation (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.