簡要描述：GS2-M小鼠胚胎干細胞培養基，ES/iPS特點
無血清、成分確定配方、無飼養層
不添加LIF也可以發揮作用
使用GSK3β和 ERK/MEK小分子抑制劑來阻斷分化

 華雅旗下紅榮微再授權代理（上海）TAKARA CellArtis，Rubicon產品 

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英文名稱：Cellartis GS2-M Mouse ES and iPS stem culture medium

中文名稱：Cellartis GS2-M小鼠胚胎干細胞和多能誘導干細胞培養基

用途：小鼠（mouse）胚胎干細胞(ES)和誘導多能干細胞(iPS)細胞的維持培養

保存：基礎培養基-20ºC；添加劑-80ºC

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

Cellartis 提供一款小鼠胚胎干細胞培養基GS2‐M，其成分確定、無血清，支持無飼養層培養，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，進而維持小鼠胚胎干細胞處于Ground state。值得一提的是，使用GS2-M培養基培養的小鼠胚胎干細胞，與添加血清和LIF的培養基培養相比，可以更好地維持小鼠胚胎干細胞處于Ground-state。

GS2-M與細胞因子leukemia inhibitory factor (LIF)一同使用時，可以通過調控Nanog信號通路實現將partial or pre‐iPS cells轉化成為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).

產品特點：

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

產品應用

小鼠部分或pre-iPS細胞轉化為*多能干細胞*

將人ES細胞或iPS細胞轉化為初始(naïve)干細胞*

維持純ES和iPS細胞的多能狀態

*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 (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 ‘naïve’ human iPS cells4.

A.對于小鼠ES和iPS細胞。

培養容器在PBS溶液（室溫下15分鐘或更長時間）中預先用0.1％明膠A型包被，或者預先用0.01％聚-L-鳥氨酸氫溴酸鹽（在37℃下30分鐘或更多）預包被， 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細胞。

請參考王偉等人（2011）更詳細的試劑和培養要求以生成和傳播“幼稚”人類iPS細胞的參考文獻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.

參考文獻：

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.

Cellartis 小鼠/大鼠胚胎干細胞培養基系列：成分確定、無血清培養基

GS2-M® 小鼠胚胎干細胞和多能誘導干細胞培養基，無飼養層，不依賴LIF

iSTEM™ 小鼠胚胎干細胞培養基，難獲得ES細胞小鼠

GS1-R® 大鼠多能干細胞培養基，需要飼養層，胚胎干細胞和胚胎生殖細胞（EG）

華雅再生醫學旗艦公司：紅榮微再（上海）生物工程技術有限公司主營細胞治療和精準醫療等產品，2018年與TAKARA繼續合作，授權代理（上海）其旗下Cellartis干細胞研究制品和Rubicon單細胞文庫構建試劑。Cellartis公司位于瑞典，擁有iPS細胞等干細胞分化為肝細胞及臟器細胞的分化誘導技術和ES細胞，ips細胞，分化細胞等干細胞相關產品。

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