GS1‐R is a compley defined, serum‐free cell culture medium that is used for the derivation, maintenance, and propagation of rat embryonic stem (ES) and embryonic germ (EG) cells. The medium contains selective small molecule inhibitors that block differentiation-inducing signals, maintaining cells in the ‘ground state’ of self-renewal and promoting cell survival and growth (1–3). Rat ES cells cultured in GS1‐R medium are germ-line competent and can be used to generate transgenic knock‐out rats using either classic homologous recombination or nuclease-based gene editing techniques.
Uses selective small molecule inhibitors to block differentiation signals from GSK3β and ERK/MEK
Culture in this medium allows rat ES cells to be efficiently derived, propagated, and genetically manipulated
產(chǎn)品應(yīng)用(ES:胚胎干細(xì)胞;EG:胚胎生殖細(xì)胞)
大鼠ES和EG細(xì)胞的分離培養(yǎng)和長期維持生殖潛力
Derivation and long‐term maintenance of germline-competent rat ES and EG cells
組份:培養(yǎng)基+添加劑
100 ml medium
100 µl 2i inhibitor supplement
自備試劑:飼養(yǎng)層細(xì)胞或預(yù)鋪培養(yǎng)容器+LIF
Rat ES and EG cells will require co-culture support from feeder cells; StemCell Sciences (UK) Ltd. recommends inactivated mouse embryonic fibroblastsseeded at 2 - 4 x 104 cells/cm2 on pre-gelatinised culture vessels.Further supplementation with an activating STAT3 signalling cytokine such as LIF is optional5.
大鼠ES和EG細(xì)胞需要來自飼養(yǎng)細(xì)胞的共培養(yǎng)支持; Stem Cell Sciences(UK)Ltd.推薦滅活的小鼠胚胎成纖維細(xì)胞接種2-4×10 4個(gè)細(xì)胞/ cm 2在pre-gelatinised的培養(yǎng)容器上。進(jìn)一步補(bǔ)充活化STAT3信號(hào)傳導(dǎo)因子如LIF。
保存:Upon receipt, store the media at -20℃ and the 2i supplement at -80℃ untilready to use.When stored under these conditions, the products are stable for12 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. Li P, et al. (2008) Germline competent embryonic stem cells derived from ratblastocysts. Cell. 135(7): 1299-1310.
2.Buehr M, et al. (2008) Capture of authentic embryonic stem cells from ratblastocysts. Cell. 135(7): 1287-1298.
3.Leitch H, et al. (2013) Naive pluripotency is associated with global DNA hypo-methylation. Nat Struct Mol Biol. 20: 311–316.
4.Tong C, et al. (2011) Generating gene knockout rats by homologous recom-bination in embryonic stem cells. Nat Protocols. 6: 827-844.
5.Blair K, et al. (2012) Culture parameters for stable expansion, genetic modi-fication and germline transmission of rat pluripotent stem cells. Biol Open.1(1): 58-65.
6.Tong C, et al. (2012) Rapid and cost-effective gene targeting in rat embryonicstem cells by TALENs. J Genet Genomics. 39(6): 275-280.