3i Stem Cell Reprogramming and Maintenance Kit
A set of three small-molecule inhibitors (CHIR-99021, SU5402 and PD184352) for the reprogramming and long-term maintenance of induced pluripotent stem cells under 3i conditions. One kit supplements up to 3 L of culture medium.
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Background & Details
The 3i Stem Cell Reprogramming and Maintenance Kit combines three well-characterized inhibitors that together hold pluripotent stem cells in a stable, self-renewing "ground state". CHIR-99021 inhibits GSK-3 and activates Wnt/β-catenin signaling. SU5402 blocks FGF receptor signaling, and PD184352 (CI-1040) blocks MEK/ERK signaling. Shutting down these differentiation-promoting pathways while supporting Wnt activity lets pluripotent cells self-renew without serum or feeder-derived factors.
Added to a suitable basal medium, the cocktail supports the reprogramming of fibroblasts into iPSCs and their maintenance afterwards. iPSCs derived under 3i conditions have been reported to show high-quality pluripotency and faster, more efficient differentiation into all three germ layers.
Each inhibitor is supplied as a separate solid. After reconstitution in DMSO, the three stocks are combined in equal volumes to give a 1000X stock that is diluted directly into medium.
Description
How to use
- Briefly centrifuge each vial before opening.
- Add sterile DMSO to each vial: CHIR-99021 1.19 mL, SU5402 1.41 mL, PD184352 4.35 mL. Vortex until fully dissolved.
- Mix equal volumes of the three stocks to obtain a 1000X 3i stock.
- Dilute 1:1000 into culture medium (final: 3 μM CHIR-99021, 0.8 μM SU5402, 0.8 μM PD184352). Sterile-filter the final medium before use.
- Store stock solutions at -20°C for up to 1 month. Aliquot to avoid repeated freeze–thaw cycles.
References
- Ying QL, et al. The ground state of embryonic stem cell self-renewal. Nature. 2008;453(7194):519-523.
- Nishihara K, et al. Induced pluripotent stem cells reprogrammed with three inhibitors show accelerated differentiation potentials with high levels of 2-cell stage marker expression. Stem Cell Reports. 2019;12(2):305-318.
Components
Each kit contains one vial of each component below (5 mg + 1 mg + 5 mg), enough to supplement up to 3 L of culture medium. The 3 Kits size contains three complete sets (up to 9 L).
| Component | Cat. No. | CAS | MW | Target | Amount | DMSO to add | Stock | Working conc. |
|---|---|---|---|---|---|---|---|---|
| Laduviglusib (CHIR-99021) | A10199 | 252917-06-9 | 465.34 | GSK-3α/β | 5 mg | 1.19 mL | 9 mM (3000X) | 3 μM |
| SU5402 | A11558 | 215543-92-3 | 296.32 | FGFR1 | 1 mg | 1.41 mL | 2.4 mM (3000X) | 0.8 μM |
| PD184352 (CI-1040) | A10212 | 212631-79-3 | 478.66 | MEK1/2 | 5 mg | 4.35 mL | 2.4 mM (3000X) | 0.8 μM |
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Specifications
| Manufacturer | Adooq Bioscience |
|---|---|
| Applications | Reprogramming of fibroblasts into induced pluripotent stem cells (iPSCs) and maintenance of pluripotent stem cells in the ground state. |
| Synonyms | 3i Cocktail; 3i Kit; CHIR-99021/SU5402/PD184352; iPSC Reprogramming Inhibitor Kit; Ground State Inhibitor Kit; CHIR99021; SU 5402; CI-1040 |