StemRNA™ 重编程试剂盒 Stemgent® StemRNATM SR Reprogramming Kit

StemRNA™ 重编程试剂盒
Stemgent® StemRNATM SR Reprogramming Kit

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

StemRNATM 重编程试剂盒StemRNA™ 重编程试剂盒                              Stemgent® StemRNATM SR Reprogramming Kit

Stemgent® StemRNATM SR Reprogramming Kit

产品概况

ReproCELL 的 Stemgent® StemRNA -SR 重编程试剂盒是细胞重编程试剂盒,结合自身复制 RNA(srRNA)和 microRNA 技术,为干细胞研究者提供安全、灵活、快速、经济、高效重编程,可将来自人外周血、脐带血、人成熟和新生成纤维细胞的内皮祖细胞(EPCs)通过重编程转化为 IPS 细胞。

该试剂盒有3种组分                                             StemRNA™ 重编程试剂盒                              Stemgent® StemRNATM SR Reprogramming Kit



● OKSiM-srRNA

● microRNA 重编程混合物

● 18R 重组蛋白

◆优势

● 重编程来自人外周血或者脐带血的 EPCs 的实验流程经过鉴定。

● 不含病毒或 DNA 的 OKSiM 重编程因子,包括自我复制 RNA 。

● 两种转染流程,免除了条件性培养基、共转染、小分子和饲养层细胞的需求。

● 25 天内制备来自 EPCs 和成纤维细胞的 iPS 细胞克隆。

● 流程可在6孔板内进行5种重编程。

血液来源

原代 EPC

建立效率

病人之间

重编程效率

外周血

18/22 = 82%

9/13 = 70%

脐带血

46/53 = 87%

33/41 = 81%

64/75 = 85%

42/54 = 78%

表 1. 来自人血液样品的 EPCs 进行有效的细胞 srRNA 重编程


来自外周血的原代 EPC 建立效率是新鲜血液和冻存的单核细胞(MNC)样品的累加。脐带血效率的数据只是来自冻存的 MNCs 。每种胶原被的 T-75 瓶最少接种 5 x 107 MNCs 。未修饰 StemRNA-SR 试剂盒步骤证明使用一个重编程实验即可得到病人之间的高效率。典型的最终结果是6孔板内每个孔有 10~20 个 iPS 细胞克隆。



◆相关产品


货号

名称

规格

00-0073

microRNA Booster Kit

microRNA增强试剂盒

1kit

00-0071

 Stemgent® mRNA Reprogramming Kit

 mRNA 重编程试剂盒

1kit

00-0075

Stemgent® StemRN -SR Reprogramming Kit

StemRNA -SR重编程试剂盒

1kit


注:本品已停产,升级版本已推出:Stemgent® StemRNA™ 3rd Generation Reprogramming Kit(又名 StemRNA-NM Reprogramming Kit,产品编号:00-0076)。

试剂盒组分

● OKSiM-srRNA, (Part No. 05-0038), 5 µg, 1 瓶

● microRNA Reprogramming Cocktail (20 µM) (Part No. 05-0036), 35 µL, 1 瓶

● B18R Recombinant Protein, Carrier-Free (Cat. No. 03-0017), 50 µg, 1 瓶

 

保存

● 保存和保质期Storage and Stability

● 所有组分均保存在 -70 °C 及其以下

● 到货后至少保存3个月

 

 

质量控制

StemRNA™-SR Reprogramming Kit 可以重编程人脐静脉内皮细胞(HUVEC)。重编程的 iPS 细胞克隆用多能干细胞标志物和细胞形态进行鉴定。所有成分均经过灭菌,不含支原体。Limited Use Licenses

Limited Use License MIT
Limited Use License iPS Academia Japan

This product also uses technology licensed from the University of California – San Diego.



推荐使用

建议使用 StemRNA-SR 制备 EPC 重编程成为 iPS.

 

● 制备 EPC 需要的材料清单-点击

● i StemRNA-SR 重编程要的材料-点击

● ISSCR 2015 (Stockholm)海报: ISSCR2015-srRNA-mobile.pdf


 

Specification Sheets

00-0075 Product Specification Sheet

Safety Data Sheets

03-0017 Safety Data Sheet

05-0036 Safety Data Sheet

05-0038 Safety Data Sheet

Product Sheets

00-0075 Product Sheet

 

[1]

Yoshioka N; Gros E; Li H-R; Kumar S; Deacon DC; Maron C; Muortri AR; Chi NC; Fu X; Yu BD; Dowdy SF. "Efficient Generation of Human iPSCs by a Synthetic Self-Replicative RNA." Cell Stem Cell 13: 246 (2013)

[2]

Geti I; Ormiston M: Touhain F; Toshner M; Movassagh M; Nichols J; Mansfield W; Southwood M; Bradley A; Rana AA; Vallier L; Morrell NW. "A practical and efficient cellular substrate for the generation of induced pluripotent stem cells." Stem Cells Trans med 1:855 (2012)

[3]

Yoshida Y; Takahashi K; Ishisaka T; Yamanaka S. "Hypoxia Enhances the Generation of Induced Pluripotent Stem Cells." Cell Stem Cell 5:237 (2009)


  

    

产品列表
产品编号 产品名称 产品规格 产品等级 备注
00-0075 Stemgent® StemRNATM SR Reprogramming Kit
StemRNATM-SR重编程试剂盒
1kit

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

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

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

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

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

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


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。

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


StemRNA 3rd Gen Reprogramming Kit

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

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

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


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

相关资料



◆视频

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

Stemgent mRNA 重编程试剂盒使用技巧

◆宣传页以及说明书


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

Stemgent_RNA Reprogramming System

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

血液-Protocol

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

尿液-Protocol

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

成纤维细胞-Protocol

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)


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