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B8 media discovery kit Qk503

B8 media discovery kit (Qk503)

For current hiPSC media comparison with B8 media to see if your cells can be maintained as efficiently while giving you the weekends off.

 

B8 hiPSC media, developed in the Burridge lab at Northwestern University, is growing in popularity thanks to its cost-effective and weekend-free cell culture credentials. B8 medium is a thermostable version of Essential 8 Medium. It is xeno-free and feeder-free medium for the growth and expansion of human pluripotent stem cells.

The B8 media Discovery kit allows the optimization of the published B8 recipe, including modified versions with higher concentrations of TGF-β1/3 (1 or 2 ng/ml), to evaluate how concentrations affect the expression of pluripotency markers. Compare B8 media with current hiPSC media to evaluate whether your cells can be maintained as efficiently with weekend-free culture.

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Kit content

  • IGF-1 (insulin-like growth factor 1)  – Qk047 – 500 µg
    Used in the maintenance of human pluripotent stem cells and is necessary for cell growth in the absence of insulin1.

  • IGF-1 LR3 (insulin-like growth factor long arginine 3) – Qk041 – 500 µg
    A synthetic analog of IGF-1. The substitutions include an Arginine substitution and an N-terminal protein extension. Consequently, IGF-1 LR3 has improved biological potency and extended half-life.

  • Human FGF2-G3 (145 aa) – Qk052 – 100 µg
    A thermostable engineered form of human FGF-2. Human FGF2-G3 145 aa comprises the 145 aa form of FGF-2 (Qk025). The functional half-life has increased from <10 h (wild-type) to >7 days (FGF2-G3).

  • Human FGF2-G3 (154 aa) – Qk053 – 100 µg
    A thermostable engineered form of human FGF-2. Human FGF2-G3 154 aa is the 154 aa mature domain of FGF-2 (Qk027). The functional half-life has increased from <10 h (wild-type) to >7 days (FGF2-G3).

  • NRG1 – Qk045 – 50 µg
    Frequently used in the maintenance of human pluripotent stem cells and has widespread use in stem cell culture media.

  • TGF-β1 PLUS – Qk010 – 25 µg
    Regulates various cellular processes, including cell proliferation, growth, differentiation, motility, and apoptosis. It is an essential growth factor in many embryonic and induced pluripotent stem cell maintenance media, including the commonly used E8, StemPro, and mTeSR media.

  • TGF beta 3 – Qk054 – 25 µg
    A member of the Transforming Growth Factor family, a family involved in regulating cell survival, proliferation and differentiation.1 TGF-β3 is used in human pluripotent stem cell maintenance medias, such as B8 media.

Quality features

  • Highly pure >98%, by SDS-PAGE quantitative densitometry

  • Highly bioactive – come with our Bioactivity Guarantee

  • Source: Expressed in E. coli 

  • Animal-free (AOF) and carrier protein-free

  • Manufactured in Cambridge, UK

  • Lyophilized

Quality assays

  • Mass spectrometry, single species with the expected mass

  • Endotoxin: <0.005 EU/μg protein (below the level of detection)

  • Recovery from stock vial: >95%

Kit description

B8 medium is specifically designed to support the growth and expansion of human pluripotent stem cells, demonstrating its ability to uphold pluripotency across various induced pluripotent stem cell (iPSC) lines as well as embryonic stem cells (ESCs) [1-2].

B8 media is a cost-effective media designed to support cell culture without the need for daily media changes. B8 media is a thermostable version of Essential 8 (E8) Medium, which is xeno-free and feeder-free, meaning it does not contain animal-derived components and does not require feeder cells for cell growth.

B8 medium provides an optimal environment for the maintenance of pluripotency in hPSCs, making it a popular choice among researchers in the field of stem cell biology and regenerative medicine. Additionally, B8 media is often used in various experimental protocols and drug discovery studies involving hPSCs.

The activation of key signaling pathways crucial for maintaining pluripotency is facilitated by specific components within the medium. Fibroblast growth factor 2 (FGF-2) and/or neuregulin 1 (NRG-1) stimulate the PI3K/AKT/mTOR and MAPK/ERK pathways by binding to FGFR1/FGFR4 or ERBB3/ERBB4 receptors, respectively. The TGF-β signaling pathway is activated by factors such as TGF-β1, nodal, or activin A, which bind to receptors TGFBR1/2 and/or ACVR2A/2B/1B/1C. Nodal activation is less common in pluripotent medium formulations due to the presence of nodal antagonists LEFTY1/2 in human pluripotent stem cells (hPSCs). The PI3K/AKT pathway, crucial for promoting cell survival and growth, is activated by insulin or insulin-like growth factor 1 (IGF-1), which bind to insulin receptor (INSR) and IGF1 receptor (IGF1R) respectively. These intricate signaling mechanisms underscore the precise formulation of this medium to sustain the delicate balance of pluripotency in human stem cells.

[1] Kuo, Hui-Hsuan et al. Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture. Stem Cell Reports, vol. 14, no. 2, 2020, pp. 256-270, doi:10.1016/j.stemcr.2019.12.007.

[2] Fonoudi, Hananeh et al. Generating a Cost-Effective, Weekend-Free Chemically Defined Human Induced Pluripotent Stem Cell (hiPSC) Culture Medium. Current Protocols in Stem Cell Biology, vol. 53, no. 1, 2020, article e110, doi:10.1002/cpsc.110.

[3] Chen, G., Gulbranson, D. R. et al. (2011). Chemically defined conditions for human iPSC derivation and culture. Nature Methods, 8(5), 424-429. doi: 10.1038/nmeth.1593

[4] Ludwig, T. E., Levenstein, M. E. et al. (2006). Derivation of human embryonic stem cells in defined conditions. Nature biotechnology, 24(2), 185-187. doi.org/10.1038/nbt1177

[5] Yao, S., Chen, S., Clark, J., Hao, E., Beattie, G. M., Hayek, A., & Ding, S. (2006). Long-term self-renewal and directed differentiation of human embryonic stem cells in chemically defined conditions. Proceedings of the National Academy of Sciences, 103(18), 6907-6912. doi: 10.1073/pnas.0602280103

[6] Guhr, A., Kobold, S., Seltmann, S., Seiler, Wullich, & Klingmüller, D. (2006). Human embryonic stem cell lines: lines from clinical grade batches. J Stem Cells Regen Med, 2, 202-213.

Additional information

Qkine is committed to manufacturing bioactive proteins of the highest quality to enhance scientific outcomes and improve reproducibility. To ensure absolute reproducibility and optimize scientific outcomes, all our products rigorously adhere to the Nine-point Qkine Quality Commitment.

Qkine products are intended solely for research purposes and ex vivo cell manufacturing. Not for therapeutic or diagnostic use.
Discovery kits are only available on orders direct from Qkine. Distributors and resellers of Qkine products are not eligible. This product is not for resale.
No additional discount promotion can be applied to the price of the Discovery Kit. Other restrictions may apply.
If you have any questions relating to this product, please email customerservice@qkine.com.

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