Abstract Number: PB0908
Meeting: ISTH 2021 Congress
Background: Several recognized protocols exist to differentiate human induced pluripotent stem cells (hiPSCs) into endothelial cells (hiPSC-ECs). Even though these hiPSC-ECs mimic primary ECs very accurately, round Weibel-Palade bodies (WPBs) and low von Willebrand factor (VWF) production indicate an immature EC phenotype of the hiPSC-ECs. However, to be used as a proper model, these cells require a mature EC phenotype. Exposure to a histone deacetylase inhibitor (HDACi) during differentiation may cause remodeling of the gene expression profile and may improve in vitro maturation of hiPSC-ECs.
Aims: To improve the maturity of the EC phenotype of hiPSC-ECs by the addition of a HDACi during and after differentiation.
Methods: Peripheral blood mononuclear cells from three healthy donors were reprogrammed into hiPSCs and subsequently differentiated into ECs. At different timepoints, the HDACi sodium butyrate was added at concentrations ranging from 0.25-2.5mM. VWF production and secretion of the hiPSC-ECs was measured, together with the expression of VWF related transcription factors.
Results: The addition of sodium butyrate did not have an effect on VWF secretion, neither at basal levels nor after histamine stimulation of the hiPSC-ECs. However, when measured in the cell lysates, a small increase in VWF production was seen, which was confirmed by confocal microscopy. Even though an increase in the number of WPBs was observed, these organelles still lacked the tubular shape and remained visible as round immature structures as shown previously in hiPSC-ECs. qPCR analysis did show an increase in the expression level of several VWF related transcription factors after the addition of sodium butyrate, leading to an increase in VWF expression and production.
Conclusions: Even though the addition of sodium butyrate increases the production of VWF in hiPSC-ECs, possibly through increased expression of transcription factors, the hiPSC-EC phenotype remains immature. However, the effects of hyperacetylation during EC differentiation and maturation needs to be investigated more.
To cite this abstract in AMA style:de Boer S, Dirven R, Laan B, Eikenboom J. Developing a Model for Studying von Willebrand Disease with hiPSC-derived Endothelial Cells [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 1). https://abstracts.isth.org/abstract/developing-a-model-for-studying-von-willebrand-disease-with-hipsc-derived-endothelial-cells/. Accessed September 25, 2021.
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