TA 01

Kinase experiment:

Compounds (TA-01) are dissolved in DMSO and tested at 10 μM concentrations against a panel of 97 kinases, which are related to stem cell differentiation and cell signaling pathways. Kinome profiling is carried out by kinase profiling service[1].

Cell experiment:

HES-3, H7 and IPS are harvested and seeded at 1.1 × 106 cells/mL as EBs in ultra-low attachment 12-well plates in bSFS medium: DMEM supplemented with 2 mM l-glutamine, 0.182 mM sodium pyruvate, 1% non-essential amino acids, 0.1 mM β-mercaptoethanol, 5.6 mg/L transferrin, 20 μg/L sodium selenite, 0.25% (w/vol) Bovine Serum Albumin and 0.25% (w/vol) Hysoy. Cells are incubated at 37°C and 5% CO2 to allow EB formation. The medium is refreshed after 1 day and then every 2-3 days. Cells are stimulated with the respective compounds (TA-01) dissolved in DMSO (1 μL DMSO/mL of media) starting from day 1 or day 4, until day 8. CHIR99021 is applied for the first 24 h only[1].

References:

[1]. Laco F, et al. Cardiomyocyte differentiation of pluripotent stem cells with SB203580 analogues correlates with Wnt pathway CK1 inhibition independent of p38 MAPK signaling. J Mol Cell Cardiol. 2015 Mar;80:56-70.