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StemRNA 3rd Gen Reprogramming Kit
StemRNA第三代重编程试剂盒

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StemRNA 3rd Gen Reprogramming Kit

StemRNA第三代重编程试剂盒 (又名StemRNA™-NM重编程试剂盒)



Stemgent RNA重编程的新产品——StemRNA第三代重编程试剂盒(旧名StemRNA-NM重编程试剂盒),结合了未经修饰的RNA和microRNA技术,使干细胞研究人员追求的多功能性、简易性和时间效益提升到一个新的层次,使人成纤维细胞,以及来自难以重编程的患者血液和尿液的细胞样品得已重编程,成为具有全能性的诱导型多功能干细胞(iPS)。



◆主要优点


● 运用灵活技术,可从多种靶细胞得到高质量的人iPS细胞系

● 即开即用。可用于来自皮肤(成纤维细胞),血液(内皮祖细胞; EPCs)和尿液(尿源性上皮细胞; UDCs)

● 细胞的重编程。


● 高效率的非整合重编程

● StemRNA-3rd Gen仅需要少至四种额外的试剂。试剂盒中的双链microRNA可增强重编程效率,提高

● iPS获得效率(成纤维细胞高达4%,EPC高达3%,UPC高达0.5%)。


● 省时的操作更快获得结果,从而提高吞吐量

● 从成纤维细胞的细胞系中得到iPS克隆需要10-14天,而从EPC或UPC中获得细胞系需要12-16天。无需筛选,

● 无需像其他技术般额外培养2-10周(3-10代)以清除载体。

 


StemRNA 3rd Gen Reprogramming Kit (00-0076)

特点

成纤维细胞

尿液 (UDCs)

血液(EPCs)

每个试剂盒可转染的孔数(6孔板)

9

3

3

转染次数

4

6-8

6-8

获得原代iPS细胞克隆所需天数

10-14

12-14

12-14

重编程效率

2-+4%

0.1-0.5%

0.4-3%

筛选

兼容异源成分的培养步骤

GMP级的RNA制备步骤



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在NutriStem™ XF / FF培养基(01-0005)中扩增后,使用StainAlive™ TRA-1-60抗体(09-0068)和DAPI对UDCs的P7原代iPS细胞进行染色


图片.png


StemRNA 3rd Gen Reprogramming Kit



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以StemRNA 3rd Gen Reprogramming Kit配合NutriStem™ XF/FF培养基(01-0005)及iMatrix-511(NP892-011)基质上培养成纤维细胞衍生的iPS细胞7代。 放大倍数:4x



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StemRNA-NM的成纤维细胞重编程s时间表



◆产品说明


规格

 1 kit

试剂盒组分

 OKSMNL NM-RNA(编号05-0040),30 µg,1 vial
 EKB NM-RNA(编号05-0041),22 µg,1 vial
 NM-microRNAs(编号05-0042),15 µg,1 vial

储存与稳定性

 1. 试剂盒三种成分均需储存在-70°C以下;
 2. 在适当的保存条件下,自到货后三个月内有效。

质量控制

 试剂盒种每种mRNA的大小与完整性均经过检测。StemRNA 3rd Gen Reprogramming
 Kit 成功进行了人成纤维细胞和人脐静脉内皮细胞RNA基础重编程功能验证。iPS细胞系完
 全重编程可通过多能性标记物的表达及正确的形态来确认。试剂盒所有组分均无菌,支原
 体检测均呈阴性。

使用许可

 该产品使用Biontech AG独家许可技术,引自专利WO 2009/088134。Biontech AG的
 WO 2007/036366,WO 2007/036366,WO 2007/036366和WO 2007/036366等多
 专利也涵盖了本产品。



◆产品列表


产品编号

产品名称

规格

00-0076

Stemgent® StemRNA™ 3rd Generation Reprogramming Kit
    (StemRNA-NM Reprogramming Kit)

1 kit



◆相关产品


 产品编号

产品名称

包装

01-0005

NutriStem XF/FF Culture Medium (500 mL)
NutriStem 无饲养层干细胞培养基

500 mL

01-0005-100

NutriStem XF/FF Culture Medium (100 mL)

NutriStem 无饲养层干细胞培养基

100 mL

03-0002

Stemfactor™ FGF-Basic, Human Recombinant
Stemfactor™ FGF-Basic,人重组

50 µg

01-0020-50

NutriFreez D10 Cryopreservation Mediu

50 mL



iMatrix-511


产品编号

产品名称

包装

NP892-011

iMatrix-511
iMatrix-511层粘连蛋白

350 μg

NP892-012

1050 μg

NP892-013

175 μg

385-07361

iMatrix-511 solution(0.5 mg/mL)
重组层粘连蛋白511-E8片段,溶液(0.5 mg/mL)

175 μg×2


iMatrix-511产品详细信息请点击:iMatrix-511层粘连蛋白


查看相关产品宣传册,请点击:Stemgent细胞重编程



参考文献


1. 

Poleganov MA; Eminli S; Beissert T; Herz S; Moon J-I; Goldmann J; Beyer A; Heck R; Burkhart I; Roldan DB; Tureci O; Yi K; Hamilton B; Sahin U. "Efficient reprogramming of human fibroblasts and blood-derived endothelial progenitor cells using nonmodified RNA for reprogramming and immune evasion." Human Gene Therapy 26:751 (2015)



Featured Publication


2.

Gagliano O; Luni C; Qui W; Bertin E; Torchio E; Galvanin S; Urciuolo A; Elvassore N. Microfluidic reprogramming to pluripotency of human somatic cells. Nat. Protocols 14:772-737 (2019).

 

The StemRNA 3rd Gen Reprogramming Kit was used to generate iPSCs from fibroblasts in a microfluidic system that used only 1500 cells in 20 µL of medium. RNA-based reprogramming is ideal for such an application since it does not require extended culture for elimination of reprogramming vectors.



Additional Publications


3.

Nakajima M; Yoshimatsu S; Sato T; Nakamura M; Okahara J; Sasaki E; Shiozawa S; Okano H. Establishment of induced pluripotent stem cells from common marmoset fibroblasts by RNA-based reprogramming. Biochem Biophys Research Commun in press:

https://doi.org/10.1016/j.bbrc.2019.05.175 (2019).


4.

Watanabe T; Yamazaki S; Yoneda N; Shinohara H; Tomioka I; Iiguchi T; Tagoto M; Ema M; Suemizu H; Kawai K; Sasaki E. Highly efficient induction of primate iPS cells by combining RNA transfection and chemical compounds. Genes to Cells in press:doi:10.1111/gtc.12702 (2019).


5.

Liu L-P; Li Y-M; Guo N-N; LI S; Ma X; Zhang Y-X; Gao Y; Huang J-L; Zheng D-X; Wang L-Y; Xu H; Hui L; Zheng Y-W. Therapeutic Potential of Patient iPSC-Derived iMelanocytes in Autologous Transplantation. Cell Reports 27:455-466.e5 (2019).


6.

Sacco AM; Belviso I; Romano V; Carfora A; Schonauer F; Nurzynska D; Montagnani S; Di Meglio F; Castaldo C. Diversity of dermal fibroblasts as major determinant of variability in cell reprogramming. J Cell Mol Med :1-13; https://doi.org/10.1111/jcmm.14316 (2019).


7.

Klein T; Klug K; Henkel L; Kwok CK; Edenhofer F; Klopocki E; Kurth I; Üceyler N. Generation of two induced pluripotent stem cell lines from skin fibroblasts of sisters carrying a c.1094C>A variation in the SCN10A gene potentially associated with small fiber neuropathy. Stem Cell Res 35:101396 (2019).


8.

Dasgupta B; Rusha E; Drukker M. iPSC generation, prime to naïve reversion & characterization and primordial germ cell differentiation of Northern White Rhino. Univ Bremen : (2019).


9.

Su S; Guntur AR; Nguyen DC; Fakory SS; Doucette CC; Leech C; Lotana H; Kelley M; Kohil J; Martino J; Sims-Lucas S; Liaw L; Vary C; Rosen CJ; Brown AC. A Renewable Source of Human Beige Adipocytes for Development of Therapies to Treat Metabolic Syndrome. Cell Reports 25:3215-3228.e9 (2018).


10.

Klein T; Henkel L; Klug K; Kwok CK; Klopocki E; Üceyler N. Generation of the human induced pluripotent stem cell line UKWNLi002-A from dermal fibroblasts of a woman with a heterozygous c.608 C>T (p.Thr203Met) mutation in exon 3 of the nerve growth factor gene potentially associated with hereditary sensory and autonomic neuropathy type 5. Stem Cell Research

https://doi.org/10.1016/j.scr.2018.10.017 (2018)



11.

Liu X; Nefzger CM; Rossello FH; Chen J; Knaupp AS; Firas J; Ford E; Pflueger J; Paynter JM; Chy HS; O'Brien CM; Huang C; Mishra K; Hodgson-Garms M; Jansz N; Williams SM; Blewitt ME; Nilsson SK; Schittenhelm RL; Laslett AL; Lister R; Polo JM. Comprehensive characterization of distinct states of human naive pluripotency generated by reprogramming. Nature Methods 14:1055 (2017)



产品编号 产品名称 产品规格 产品等级 产品价格
00-0076 StemRNA™-NM Reprogramming Kit
StemgentRNA™-NM重编程试剂盒
1 kit
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