Cell lines

CHO-IR cells or primary rat hepatocytes

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below - 20 ℃ for several months.

Reacting condition

0.01 ~ 10 μM

Applications

In CHO-IR cells or primary rat hepatocytes, CHIR-99021 dose-dependently resulted in a two- to three-fold stimulation of the GS activity ratio above basal. In CHO-IR cells, CHIR-99021 induced GS activation with the EC50 value of 0.763 μM.

Animal models

ZDF rats

Dosage form

16 or 48 mg/kg; p.o.

Applications

In ZDF rats, CHIR-99021 treatment (16 or 48 mg/kg; p.o.; 1 hr before oral glucose challenges) significantly improved glucose tolerance with 14% and 33% reduction in plasma glucose, respectively. Moreover, CHIR-99021 at a higher dose also reduced hyperglycemia before the oral glucose challenge.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

CHIR-99021 is a highly specific glycogen synthase kinase-3 (GSK-3) inhibitor which can inhibit both isoforms with IC50 of 10 nM (GSK-3α) and 6.7 nM (GSK-3β).

CHIR-99021 was proved to promote self-renewal and maintain pluripotency of both B6 and BALB/c ES cells via stabilizing the downstream effectors like c-Myc and β -catenin[1]. In J1 mESC cells, CHIR-99021 played an important role in maintaining the colony morphology as well as the self-renewal when combined with leukemia inhibitory factor (LIF). CHIR-99021 has shown to regulate multiple signaling pathways which involve Wnt/β-catenin, TGF-β, Nodal and MAPK, and the expression of epigenetic regulatory genes like Dnmt3l[2].

CHIR-99021 has been demonstrated to promote DN3 thymocytes proliferation and differentiation in the absence of pre-TCR signaling, Notch1 signaling or CXCL12[3]. However, study has also found that higher concentration (10 μM but not 1 μM or 3 μM) of CHIR99021 might selectively inhibit differentiation by activating IL-7 signaling pathway[3].

Reference:
1.  Ye S1, Tan L, Yang R, Fang B, Qu S, Schulze EN, Song H, Ying Q, Li P. Pleiotropy of glycogen synthase kinase-3 inhibition by CHIR99021 promotes self-renewal of embryonic stem cells from refractory mouse strains. PLoS One. 2012;7(4):e35892.
2.  Wu Y1, Ai Z, Yao K, Cao L, Du J, Shi X, Guo Z, Zhang Y. CHIR99021 promotes self-renewal of mouse embryonic stem cells by modulation of protein-encoding gene and long intergenic non-coding RNA expression. Exp Cell Res. 2013
3.  PLoS One. 2013;8(3):e58501. doi: 10.1371/journal.pone.0058501. Epub 2013 Mar 20. Schroeder JH1, Bell LS, Janas ML, Turner M. Pharmacological inhibition of glycogen synthase kinase 3 regulates T cell development in vitro. PLoS One. 2013;8(3):e58501.