Cell experiment:

The differentiation of iPS cells to cardiomyocytes (CM) is induced by embryoid body (EB) formation. When iPS cells reached 70% confluency in 10-cm dishes, cells are digested using 0.25% trypsin/EDTA. Cell pellets are re-suspended in differentiation medium (DMEM with 20% FBS and 10 ng/mL BMP4) to a final concentration of 200,000 cells/mL. Cell suspensions are added to 6-well plates with Ulta-Low Attachment surfaces for 4 d to initiate EB formation. On day 5, EBs are cultured on 0.1% gelatin-coated dishes for 14 d using CF culture medium for the outgrowth of cardiac structures. At this stage, iPS cells undergoing EB formation are termed iPS derivates (iPSD)[1].

NSC 14613 is a pluripotent cell-specific inhibitor (PluriSIn) that selectively eliminates human pluripotent stem cells (hPSCs) leaving a large array of progenitor and differentiated cells unaffected. Among 15 identified PluriSIns, NSC 14613 (also known as PluriSIn #1) is a derivative of N-acyl phenylhydrazine that shares a common structure moiety, phenylhydrazine (Ph-N[H,C]-NH) with 9 other PluriSIns. NSC 14613 is also an inhibitor of stearoyl-coA desaturase (SCD1) that inhibits the activity of SCD1 in hPSCs causing the imbalance between substrates and products and subsequently resulting in the accumulation of palmitate and stearate and the deprivation of oleate. Study results have suggested that NSC 14613 is able to induce ER stress, protein synthesis attenuation and apoptosis in hPSCs and can be used to prevent teratoma formation from tumorigenic undifferentiated cells.

Reference

[1].Ben-David U, Gan QF, Golan-Lev T, Arora P, Yanuka O, Oren YS, Leikin-Frenkel A, Graf M, Garippa R, Boehringer M, Gromo G, Benvenisty N. Selective elimination of human pluripotent stem cells by an oleate synthesis inhibitor discovered in a high-throughput screen. Cell Stem Cell. 2013 Feb 7;12(2):167-79