Can Differentiation of iPSC-Derived Cardiomyocytes be Improved by Culture on Rationally-Selected Extracellular Matrix Proteins

Barnes, Ashlynn

Undergraduate Honors Thesis

Can Differentiation of iPSC-Derived Cardiomyocytes be Improved by Culture on Rationally-Selected Extracellular Matrix Proteins 公开 Deposited

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Citeable URL: https://scholar.colorado.edu/concern/undergraduate_honors_theses/qz20st97w

Abstract

Current protocols used to differentiate human induced pluripotent stem cells (iPSCs) into cardiomyocytes (CMs) are lengthy and produce immature CMs. Additionally, current standard protocols use Matrigel, which is not a defined substrate. To investigate whether alternative substrates could increase efficiency of iPSC-CM differentiation protocols, iPSC-CMs were grown on seven rationally-selected extracellular matrix (ECM) proteins. Studies comparing iPSC-CM differentiation on multiple substrates compared to Matrigel have been limited. We hypothesized iPSC-CM differentiation speed and maturity could be improved by differentiation on rationally-selected ECM proteins. To test this hypothesis, the speed at which the iPSC-CMs differentiated as wells as maturity of the resultant iPSC-CM were tested. To analyze speed, confluency over time and days until contractility onset were quantified. To analyze maturity of the cells, expression of maturation marker MYL7 was monitored. The iPSC-CMs differentiated on Laminin-521, Laminin-221 + 521, and Laminin-332 showed increased confluency during differentiation compared to the Matrigel control. The date of contractions onset observed of the iPSC-CMs grown on ECM proteins Laminin-521, and Laminin-221 + 521 was earlier compared to Matrigel. Laminin-521, Laminin-221 + 521, mouse Laminin-111, and Laminin-332 ECM proteins had less variability in date of contraction onset compared to Matrigel. The iPSC-CMs cultured on Laminin-521 and Laminin-221 + 521 were observed to contract earlier, have less variability in day of differentiation contraction onset, and grow to a higher confluency quicker. Culturing iPSCs on select ECM proteins can result in quicker iPSC-CM differentiations.

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