ID-8 is a highly potent and selective small molecule inhibitor of dual-specificity tyrosine phosphorylation-regulated kinase (DYRK).  It could support Wnt-induced human embryonic stem cell proliferation and survival without differentiation. Using ID-8, a novel and simple chemically defined xeno-free culture system was developed that allows for long-term expansion of human pluripotent stem cells without FGF or TGF-beta pathway activation. These culture conditions do not include xenobiotic supplements, serum, serum replacement, or albumin. Using this culture system, several human pluripotent cell lines maintained pluripotency (>20 passages) and a normal karyotype and still retained the ability to differentiate into derivatives of all three germ layers. This Wnt-dependent culture system should provide a platform for complete replacement of growth factors with chemical compounds.

How to Use: 

• In vitro:  ID-8 was used at 0.5 µM final concentration in cell culture.
• In vivo:  n/a

Reference:

• 1. Hasegawa K, et al. Wnt signaling orchestration with a small molecule DYRK inhibitor provides long-term xeno-free human pluripotent cell expansion. (2012)  Stem Cells Transl Med. 1(1):18-28.
• 2. Miyabayashi T, et al. Indole derivatives sustain embryonic stem cell self-renewal in long-term culture. (2008) Biosci Biotechnol Biochem. 72(5):1242-8.
• 3. Firestone AJ, et al. Controlling destiny through chemistry: small-molecule regulators of cell fate. (2010) ACS Chem Biol. 5(1):15-34.

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