Feeder-independent culture system reduces inherent variability, cost, and burden of feeder preparation
Serum-free formulation contains pre-screened quality components for reproducible results
Facilitates highly efficient reversion to naïve-like state with stable domed morphology, naïve gene expression profiles, and low levels of spontaneous differentiation without the need for exogenous genes
Maintains naïve-like pluripotency without inclusion of bFGF
Revert primed human embryonic (ES) and induced pluripotent stem (iPS) cells to a naïve-like state and maintain cells in a naïve-like state under hypoxic conditions without bFGF or feeder cells.
Developed under license from the Weizmann Institute of Science, this serum-free cell culture medium produces robust cultures with features of a naïve-like state, such as tightly packed, domed colonies with refractive edges. Human ES/iPS cells cultured in RSeT™ Feeder-Free Medium show increased expression of key transcripts associated with the naïve-like state, such as KLF17, KLF2, KLF4, and TFCP2L1, and can either be differentiated or converted back to a primed state by culture in mTeSR™1 and then differentiated.
Figure 1. Schematic of Reversion of Primed to Naïve-Like hPSCs with RSeT™ Feeder-Free
Primed hPSCs are plated as aggregates in mTeSR™1. On day 1, mTeSR™1 is replaced with RSeT™ Feeder-Free, the cultures are transferred to hypoxic conditions, and the medium is exchanged every other day. By day 4 or 5, the colonies are generally large enough to be passaged. During the initial culture in RSeT™ Feeder-Free, colonies expand and begin to adopt a tightly-packed, highly domed morphology characteristic of naïve-like stem cells with smooth and refractive colony edges as early as passage 1.
Figure 2. hPSCs Maintained in RSeT™ Feeder-Free are Reverted to a Naïve-Like State and Express High Levels of Naïve-Associated Genes
(A) A representative image of hPSCs that reverted to a naïve-like state after being cultured in RSeT™ Feeder-Free for 1 passage. During reversion, colonies change from a flat morphology to a domed morphology characteristic of naïve-state hPSCs. (B) Expression of naïve-associated genes (KLF2, KLF4, KLF17, TFCP2L1, STELLA, and DNMT3L) in hPSCs that were reverted to a naïve-like state in RSeT™ Feeder-Free. Expression levels were measured by qPCR and normalized to levels in primed hPSCs.
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
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