RSeT™ Medium (2-component)
Defined medium for naïve-like human pluripotent stem cells
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Overview
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).
Data Figures
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.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Resources and Publications
Educational Materials (9)
Legal Statement:
Yeda Research and Development Ltd., an Israeli corporation (“YEDA”) is the owner of all right and title to certain patents applications and other proprietary rights which cover (i) the use of the product you are purchasing, such product comprising of media culture compositions to produce novel fully Naïve Induced Pluripotent Stem Cells (“iPSCs”) (“the Product”) and (ii) the use of the iPSCs themselves. By purchase of the Product, you shall have, subject to the terms of use below, a limited non-exclusive licence to use the Products to generate iPSCs and the iPSCs themselves. YOU HEREBY AGREE TO USE THE PRODUCT AND ANY RESULTS OR PHYSICAL MATTER, INCLUDING IPSCS, GENERATED FROM THE USE OF THE PRODUCT, OR ANY COPIES OR DERIVATIVES OF ANY OF THE FOREGOING, SOLELY FOR THE PURPOSE OF CONDUCTING NON-COMMERCIAL RESEARCH ACTIVITIES AND PUBLISHING THE RESULTS OF SUCH ACTIVITIES. YOU HEREBY ACKNOWLEDGE THAT ANY USE THEREOF FOR PURPOSES OTHER THAN NON-COMMERCIAL RESEARCH AND PUBLICATION IS PROHIBITED AND SHALL BE PROSECUTED TO THE FULL EXTENT OF THE LAW. IN THE EVENT YOU WISH TO USE THE ABOVE FOR ANY OTHER PURPOSE, A SEPARATE LICENSE IS REQUIRED. YOU CAN CONTACT YEDA AT INFO.YEDA@WEIZMANN.AC.IL FOR THE PURPOSE OF OBTAINING A SEPARATE LICENCE IN THIS REGARD, YEDA BEING IN NO WAY OBLIGATED TO ENTER INTO OR NEGOTIATE SUCH A LICENCE WITH YOU. Nothing contained herein shall be deemed to be a representation or warranty, express or implied, by YEDA that patent applications relating to the Product and the iPSCs will be granted; (ii) that patents obtained on any of the said patent applications are or will be valid or will afford proper protection; (iii) that the use of the Product or the iPSCs will not infringe the rights of any third party; or (iv) as to the potential, use, exploitability, merchantability, suitability, effectiveness, fitness for purpose or safety of the Product and the iPSCs. Under no circumstances shall YEDA be liable for any claims, demands, liabilities, costs, losses, damages or expenses (including legal costs and attorney's fees) of whatever kind (any of the above, a “Loss”) caused to or suffered by any person or entity that directly or indirectly arise out of the use of the Product or the iPSCs.
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
