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.
產(chǎn)品特點
成分*確定的配方;無血清
利用選擇性小分子抑制劑阻斷來自GSK3β和ERK / MEK的分化信號
在該培養(yǎng)基中培養(yǎng)可以有效地擴增大鼠ES細胞并進行遺傳操作
Compley defined formulation; serum-free
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)品應用(ES:胚胎干細胞;EG:胚胎生殖細胞)
大鼠ES和EG細胞的分離培養(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)層細胞或預鋪培養(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細胞需要來自飼養(yǎng)細胞的共培養(yǎng)支持; Stem Cell Sciences(UK)Ltd.推薦滅活的小鼠胚胎成纖維細胞接種2-4×10 4個細胞/ cm 2在pre-gelatinised的培養(yǎng)容器上。進一步補充活化STAT3信號傳導因子如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:
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.