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GS2-M小鼠胚胎干細(xì)胞培養(yǎng)基，ES/iPS特點
無血清、成分確定配方、無飼養(yǎng)層
不添加LIF也可以發(fā)揮作用
使用GSK3β和 ERK/MEK小分子抑制劑來阻斷分化
 

英文名稱：Cellartis GS2-M Mouse ES and iPS stem culture medium
中文名稱：Cellartis GS2-M小鼠胚胎干細(xì)胞和多能誘導(dǎo)干細(xì)胞培養(yǎng)基
用途：小鼠（mouse）胚胎干細(xì)胞(ES)和誘導(dǎo)多能干細(xì)胞(iPS)細(xì)胞的維持培養(yǎng)
保存：基礎(chǔ)培養(yǎng)基-20oC；添加劑-80oC
 

小鼠胚胎干細(xì)胞是*被體外分離培養(yǎng)并建系的干細(xì)胞，目前其相關(guān)研究已經(jīng)比較完善并投入到了實際應(yīng)用中，很多其他種屬的干細(xì)胞研究也都會參考小鼠胚胎干細(xì)胞的研究成果。一個好的小鼠胚胎干細(xì)胞體外培養(yǎng)體系，需要同時滿足操作簡便、結(jié)果穩(wěn)定、增長迅速、可重復(fù)性好、有效維持胚胎干細(xì)胞處于Ground-state等特點。

Cellartis 提供一款小鼠胚胎干細(xì)胞培養(yǎng)基GS2‐M，其成分確定、無血清，支持無飼養(yǎng)層培養(yǎng)，通過添加兩種特別的小分子抑制劑（CHIR99021，PD0325901=2i）來阻斷分化信號通路GSK3β和 ERK/MEK，進(jìn)而維持小鼠胚胎干細(xì)胞處于Ground state。值得一提的是，使用GS2-M培養(yǎng)基培養(yǎng)的小鼠胚胎干細(xì)胞，與添加血清和LIF的培養(yǎng)基培養(yǎng)相比，可以更好地維持小鼠胚胎干細(xì)胞處于Ground-state。


GS2-M與細(xì)胞因子leukemia inhibitory factor (LIF)一同使用時，可以通過調(diào)控Nanog信號通路實現(xiàn)將partial or pre‐iPS cells轉(zhuǎn)化成為fully pluripotent iPS cells(1, 2)。

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).


產(chǎn)品特點：
特點
優(yōu)勢
    由Dr.Austin Smith研發(fā)
    在很多雜志上發(fā)表，實驗數(shù)據(jù)可信，可放心使用
    成分確定、無血清
    批次間*，無需擔(dān)心批次間差異
    無飼養(yǎng)層培養(yǎng)
    不需要準(zhǔn)備飼養(yǎng)層細(xì)胞，
    節(jié)省了準(zhǔn)備飼養(yǎng)層細(xì)胞的時間和精力
    使用GSK3β和 ERK/MEK抑制劑
    維持細(xì)胞處于強的Ground state，
    有效制備嵌合體小鼠，形成生殖系傳遞。
    不添加LIF也可以發(fā)揮作用
    組分簡單，不需要額外購買LIF，
    更易于準(zhǔn)備完全培養(yǎng)基，節(jié)省了LIF的費用

 

Compley defined, serum-free formulation

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)化為完全多能干細(xì)胞*

將人ES細(xì)胞或iPS細(xì)胞轉(zhuǎn)化為初始(na?ve)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 na?ve 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 (at room temperature for 15 minutes or more), or precoated sequentially with 0.01% poly-L-ornithine hydrobromide (at 37℃ for 30 minutes or more), washing twice with 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 ‘na?ve’ human iPS cells4.

A.對于小鼠ES和iPS細(xì)胞。

培養(yǎng)容器在PBS溶液（室溫下15分鐘或更長時間）中預(yù)先用0.1％明膠A型包被，或者預(yù)先用0.01％聚-L-鳥氨酸氫溴酸鹽（在37℃下30分鐘或更多）預(yù)包被， 用PBS洗滌2次，然后用PBS中的10μg/ mL層粘連蛋白（37℃下3小時以上）洗滌。

B.對于人類iPS細(xì)胞。

請參考王偉等人（2011）更詳細(xì)的試劑和培養(yǎng)要求以生成和傳播“幼稚”人類iPS細(xì)胞的參考文獻(xiàn)4。


保存：Upon receipt, store the media at -20℃ and the GS2-M 2i inhibitor supplement at -80℃ until ready to use. When stored under these conditions, the products are 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.