GS2‐M is a compley defined, serum‐free cell culture media formulation for the generation and long-term maintenance of human and mouse embryonic stem (ES) and induced pluripotent stem (iPS) cell lines. The medium contains two selective small molecule inhibitors that block differentiation‐inducing signals and promote cell survival.
GS2-M medium can be used in combination with the self-renewal cytokine leukemia inhibitory factor (LIF) to convert partial or pre‐iPS cells from mouse into fully pluripotent iPS cells via upregulation of Nanog (1, 2).
Uses selective small molecule inhibitors to block differentiation signals from GSK3β and ERK/MEK and maintain ES and iPS cells in a pluripotent state
Allows long-term maintenance of germline-competent mouse and human ES and iPS cells
產(chǎn)品應(yīng)用
小鼠部分或pre-iPS細(xì)胞轉(zhuǎn)化為*多能干細(xì)胞*
將人ES細(xì)胞或iPS細(xì)胞轉(zhuǎn)化為初始(naïve)干細(xì)胞*
維持純ES和iPS細(xì)胞的多能狀態(tài)
*在Forskolin和/或LIF存在下
Conversion of partial or pre-iPS cells from mouse into fully pluripotent stem cells*
Conversion of human pre-ES or iPS cells into naïve stem cells*
Maintenance of pure ES and iPS cells in a pluripotent state
*In the presence of Forskolin and/or LIF
組份:
100 ml medium
100 µl 2i inhibitor supplement
自備試劑:
A. For mouse ES and iPS cells.
Culture vessels are pre-coated with either 0.1% gelatin type A in PBS solution (atroom temperature for 15 minutes or more), or precoated sequentially with 0.01%poly-L-ornithine hydrobromide (at 37℃ for 30 minutes or more), washing twicewith PBS, followed by 10 μg/mL laminin in PBS (at 37℃ for 3 hours or more).
B. For human iPS cells.
Please refer to Wang W, et al. (2011) reference for more detailed reagent and culture requirements to generate and propagate ‘naïve’ human iPS cells4.
保存:Upon receipt, store the media at -20℃ and the GS2-M 2i inhibitor supplementat -80℃ until ready to use. When stored under these conditions, the productsare stable for 12 months from the date of manufacture (see label).Once thawed and combined, store at 4℃ and use within 2 weeks.This product is light sensitive, and should be protected from light.
參考文獻(xiàn):
1. Silva J, et al. (2008) Promotion of reprogramming to ground state pluripo-tency by signal inhibition. PLoS Biol. 6: e253.
2.Silva J, et al. (2009) Nanog is the gateway to the pluripotent ground state.Cell. 138(4): 722-737.
3.Nichols J, et al. (2009) Validated germline-competent embryonic stem cell lines from non-obese diabetic mice. Nat Med. 15: 814-818.
4.Wang W, et al. (2011) Rapid and efficient reprogramming of somatic cells to induced pluripotent stem cells by retinoic acid receptor gamma and liver receptor homolog. PNAS (USA). 108(45): 18283-18288.
5.Reviewed in: Malaver-Ortega LF, et al. (2012) Theriogenology. 78(8): 1749-1762.