Effect of stearoyl-CoA desaturase-1 inhibition during differentiation of human induced pluripotent stem cells on endodermic markers expression and global protein palmitoylation

Abstract

Stearoyl-CoA desaturase 1 (SCD1) is required for de novo synthesis of fatty acid in stem cells. Methods: The hiPSCs were cultured in an endoderm inducing medium containing activin A and low defined fetal bovine serum for 4 days in the presence of an SCD1-specific inhibitor on days 0, 1 and 2. RT-PCR was used for evaluation of the stemness endoderm markers. The yield of three germ layers endoderm, mesoderm and ectoderm were assessed using flow cytometry. In addition, cell proliferation was measured by ELISA, cell cycle assay was performed using flow cytometry and total protein acylation was assessed using click reaction. Results: SCD1 inhibition showed no significant change in cell proliferation on none of the treatment days. The population of cells with endodermic features decreased at the end of differentiation when SCD1 was inhibited at the day 0 and effectively rescued by oleate supplementation. Moreover, SCD1 inhibition at day 0 preserved hiPSCs stem cell properties without a shift toward mesodermal or ectodermal differentiation. Only, treatment of cells with SCD1 inhibitor at days 0 decreased fluorescent intensity of total protein acylation and increased S phase length. Treatment at two subsequent days of endoderm induction had no significant effect on endoderm specific markers, fluorescent intensity and S phase.