Undergraduate Honors Thesis


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

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  • 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.

Date Awarded
  • 2022-03-31
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Last Modified
  • 2022-05-25
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