RSeT™ Medium (2-component)

Defined medium for naïve-like human pluripotent stem cells

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RSeT™ Medium (2-Component)

Defined medium for naïve-like human pluripotent stem cells

500 mL Kit
Catalog #05978
349 USD

Overview

RSeT™ Medium is a defined cell culture medium used for the reversion of primed human pluripotent stem cells (hPSCs) to a naïve-like state and maintenance of naïve-like hPSCs under feeder-dependent and hypoxic conditions. RSeT™ Medium contains pre-screened quality components and does not contain bFGF or TGFβ. It is compatible with human embryonic stem (ES) cells and human induced pluripotent stem (iPS) cells.
hPSCs cultured in RSeT™ Medium exhibit features of a naïve-like state such as tightly packed, domed colonies with refractive edges. Key transcripts associated with naïve-like hPSCs such as KLF2, KLF4, and TFCP2L1 show increased expression in hPSCs cultured in RSeT™ Medium. RSeT™ hPSCs can be converted back to a primed state by culture in mTeSR™1 and can then be differentiated with STEMdiff™ products such as STEMdiff™ Definitive Endoderm Kit (Catalog #05110), STEMdiff™ SMADi Neural Induction Kit (Catalog #08581), and STEMdiff™ Mesoderm Induction Medium (Catalog #05220).
Advantages:
• Easily passage as single cells while maintaining normal karyotype
• Maintains naïve-like pluripotency without bFGF or TGFβ
• Defined medium contains pre-screened quality components for reproducible results
• No exogenous genes required for reversion to naïve-like state
Components:
  • RSeT™ Basal Medium, 400 mL
  • RSeT™ 5X Supplement, 100 mL

Scientific Resources

Educational Materials

(9)

Data and Publications

Data

Figure 1. Schematic of Reversion of Primed to Naïve-Like hPSCs

Primed hPSCs are plated as aggregates in mTeSR™1 onto irradiated mouse embryonic fibroblasts (iMEFs). On day 1, mTeSR™1 is replaced with RSeT™, and the medium is exchanged daily. By day 5 or 6, the colonies are generally large enough to be passaged. During the initial culture in RSeT™ medium, colonies expand and begin to adopt domed shape characteristic of naïve-like stem cells and can continue to be propagated in RSeT™.

Figure 2. ES and iPS Cells Can Be Reverted to a Naïve-Like State

Representative images of human (A) ES cells (H1) and (B) iPS cells (STiPS-F016) lines that reverted to a naïve-like state after cultured in RSeT™ for 6 and 10 passages, respectively. (C) During reversion, colonies change from a flat morphology to a domed morphology characteristic of naïve-state hPSCs. Once naïve-like cultures are established, typically > 80% of colonies have a dome-shaped morphology.

Figure 3. ES and iPS Cells Can Be Converted Back into a Primed State

Representative images of human ES cells (H1) cultured in (A) RSeT™ for 3 passages and then (B) reconverted into a primed state in mTeSR™1 for 5 days.

Figure 4. hPSCs Maintained in RSeT™ Express High Levels of Factors Associated with Naïve-Like PSCs

Expression of markers associated with naïve-like PSCs (DNMT3L, KLF17, KLF2, KLF4, NANOG, TBX3, and TCFP2L1) in (A) WLS-1C and (B) STiPS-F019 iPS cells that were reverted back to a naïve-like state after culturing in RSeT™. Expression levels were measured by quantitative PCR (qPCR) and normalized to levels in primed STiPS-F019 iPS cells.

Figure 5. hPSCs Maintained in RSeT™ Display Normal Karyotype after Long-Term Passaging

Representative karyograms of (A) ES cells (H1) and (B) iPS cells (WLS-1C) lines that were cultured in RSeT™ medium for 14 and 15 passages, respectively.

STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.