标签归档：reprogramming

Self-replicative RNA Reprogramming kit

发表于由

Self-replicative RNA Reprogramming kit


Stemgent,  ReproCELL集团成员，发布了卓越表现的自我复制RNA重编程试剂盒


YOKOHAMA, 日本–(BUSINESS WIRE)—我们非常高兴的宣布Stemgent Inc. (Cambridge, MA), ReproCELL 集团公司(Yokohama, Japan – JASDAQ: 4978) 发布了新 Stemgent StemRNA™-SR 重编程试剂盒。这是一款可用于血液细胞的重编程的商业化RNA 重编程试剂盒。非病毒，非DNA的重编程试剂盒和自我复制RNA（srRNA）和microRNA技术配套使用，使干细胞研究者更安全，更灵活，更经济，更高效的进行细胞重编程。也可用于人成纤维细胞和血液来源的内皮祖细胞（EPCs）重编程。EPCs基因稳定，可从新鲜和冷冻保存人外周血和脐带血有效建立。该项技术在瑞典斯德哥尔摩举办的的干细胞研究国际学会(ISSCR)第13届年会议时介绍。

“采用非整合RNA重编程技术的Stemgent StemRNA-SR处理病人血液样品，对血液细胞进行重编程是研发临床相关的iPS细胞系重要进步。Joseph Gentile (Chief Executive Officer, Stemgent).

 

关于 Stemgent ( ReproCELL 集团公司

Stemgent与世界上优秀的干细胞科学家一起研发干细胞研究领域的革新技术和运用。他的使命是简化和支持重编程 Stemgent干细胞试剂提供给全世界的客户。

◆相关阅读


细胞重编程——指南目录

 

1.    什么是细胞重编程

2.    细胞重编程概述

3.    mRNA重编程

4.    病毒重编程

5.    产品列表 



◆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

StemRNATM-SR重编程试剂盒

1kit

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

发表于由

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

否

否

否

兼容异源成分的培养步骤

是

否

否

GMP级的RNA制备步骤

是

是

是

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 － －

Stemgent® mRNA 重编程试剂盒 Stemgent® mRNA Reprogramming Kit

发表于由

Stemgent® mRNA 重编程试剂盒
Stemgent® mRNA Reprogramming Kit

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

Stemgent® mRNA 重编程试剂盒Stemgent® mRNA 重编程试剂盒 Stemgent® mRNA Reprogramming Kit

Stemgent® mRNA Reprogramming Kit

◆产品概况

Stemgent® mRNA 重编程试剂盒与 microRNA Booster 试剂盒（00-0073）配套使用可以迅速，安全，有效制备人 iPS 细胞。该方法不发生整合，不含病毒，与临床研究相关。与慢病毒，仙台病毒和其他方法相比，16天内制备 iPS 细胞，３周内细胞可进行传代培养。慢病毒，仙台病毒和其他方法还需要花20周时间确认没有病毒残留。该系统完全免除了病毒相关的生物污染问题的顾虑。

 Stemgent® mRNA 重编程试剂盒包括５种重编程因子，监测转染效率的细胞核 GFP 标记蛋白，Stemgent Pluriton™ 重编程培养基， B18R 蛋白。试剂盒和操作方法都经过充分的检测和验证。该试剂盒已经能够将人 BJ 成纤维细胞进行重编程，该细胞可作为对照。Stemgent® mRNA 重编程试剂盒可以使研究正更容易调控蛋白表达水平的时间，量化控制重编程因子。



◆优势

●　非整合-无需进行基因组整合

●　快速重编程，至少 12 天制备克隆

●　量化控制各种重编程因子

●　不需要进行多次传代和细胞克隆筛选

●　在正常细胞和疾病细胞上进行验证

●　提供培训课程（收费）和在线支持

Stemgent® mRNA 重编程试剂盒 Stemgent® mRNA Reprogramming Kit

Stemgent® mRNA 重编程试剂盒 Stemgent® mRNA Reprogramming Kit


 使用 mRNA 重编程系统制备 iPS 时程图片。在第 6-10 天观察到的 

 iPS细胞形态（放大 10×）。


 使用 mRNA 重编程系统制备 iPS 时程图片，第 12 天和第 14 天。

 第14天用 TRA-1-60 荧光染色（10×）。

◆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

StemRNATM -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）。

视频

Stemgent® mRNA 重编程试剂盒 Stemgent® mRNA Reprogramming Kit

Stemgent mRNA 重编程试剂盒使用技巧

Kit Contents

●    2 vials – Stemgent Oct4 mRNA, Human (Cat. No. 05-0014), 20µg

●    1 vial – Stemgent Klf4 mRNA, Human (Cat. No. 05-0015), 20µg

●    1 vial – Stemgent Sox2, Human(Cat. No. 05-0016), 20µg

●    1 vial – Stemgent Lin28 mRNA, Human(Cat. No. 05-0017), 20µg

●    1 vial – Stemgent c-Myc mRNA, Human(Cat. No. 05-0018), 20µg

●    1 vial – Stemgent nGFP mRNA (Cat. No. 05-0019), 20µg

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

●    0.2 ml of Pluriton™ Supplement 2500X (Cat. No. 01-0016)

●    500 ml of Pluriton™ Medium (Cat. No. 01-0015)

 

Storage and Stability

1.      Store the box (Part 1 of 2) containing the 7 vials of mRNA, the 1 vial of B18R, and the 1 vial of Pluriton Supplement at or below -70°C. 

2.      Store the bottle (Part 2 of 2) of Pluriton Medium at -20°C.

3.    All kit components are stable for a minimum of 3 months from date of receipt, when stored as directed.

 

Quality Control

The Stemgent mRNA Reprogramming Kit is functionally tested for successful mRNA-based reprogramming of BJ human fibroblasts. Complete reprogramming of iPS cell colonies is confirmed by continued expression of pluripotency markers and expansion of primary iPS colonies into stable lines. All components are sterile and have tested negative for mycoplasma.


Recommended Usage

Refer to the Stemgent Protocol “Reprogramming Human Fibroblasts using the Stemgent mRNA Reprogramming Kit” for detailed instructions on material handling and productive use of the Stemgent mRNA Reprogramming Kit.


Documents

Limited Use License MIT

Limited Use License iPS Academia Japan


Related Products

●   Stemgent Oct4 mRNA, Human (Cat. No. 05-0014)

●   Stemgent Klf4 mRNA, Human (Cat. No. 05-0015)

●   Stemgent Sox2 mRNA, Human(Cat. No. 05-0016)

●   Stemgent Lin-28 mRNA, Human(Cat. No. 05-0017)

●   Stemgent c-Myc mRNA, Human(Cat. No. 05-0018)

●   Stemgent nGFP mRNA (Cat. No. 05-0019)

●   Stemedia™ NutriStem™ XF/FF Culture Medium (Cat. No. 01-0005)

●   Stemolecule™ Y27632 (Cat. No. 04-0012)

●   Stemfactor™ Fibroblast Growth Factor-basic (Human Recombinant) (Cat. No. 03-0002)

●   microRNA Booster Kit (00-0073)

      ●   RNA Transfection Kit (00-0069)

      ●   mRNA Reprogramming Factors Set: hOKSML (00-0067)

      ●   NuFF-RQ™ IRR (GSC3002G)

●   FGF-basic, Human Recombinant (03-0002)

●   XF/FF Culture Medium (01-0005)

      ●   TRA-1-60 Antibody (DyLight™ 488), Mouse anti-Human (09-0068)

      ●   TRA-1-81 Antibody (DyLight™ 488), Mouse anti-Human (09-0069)

      ●   TRA-1-81 Antibody (Purified), Mouse anti-Human (09-0011)

      ●   TRA-1-60 Antibody (Purified), Mouse anti-Human (09-0010)

●   Nanog Antibody (Affinity Purified), Rabbit anti-Mouse/Human (09-0020)

●   Oct4 Antibody (Affinity Purified), Rabbit anti-Mouse/Human (09-0023)

●   SSEA-4 Antibody (Purified), Mouse anti-Human (09-0006)

●   StemRNA™-SR Reprogramming Kit (00-0075)


Specification Sheets

●    00-0071 Product Specification Sheet

Safety Data Sheets

●   03-0017 Safety Data Sheet

●   00-0071 Safety Data Sheet

●   01-0015 Safety Data Sheet

●   05-XXXX mRNA Safety Data Sheet

●   01-0016 Safety Data Sheet

Manuals

●   User Manual: Stemgent mRNA Reprogramming System (updated January 2012)

Application Notes

●   Generation of Safe, Integration-free Human Induced Pluripotent Stem (iPS) Cells using the mRNA Reprogramming System

Brochures

●   Brochure: Stemgent mRNA Reprogramming System

Other Documents

●   Scientific Poster: mRNA Reprogramming SLAS 2011

●   FAQs: mRNA Reprogramming

●   Limited Use License MIT

●   Limited Use License iPS Academia Japan

Product Sheets

●   mRNA Reprogramming System: The fastest and safest non-integrating reprogramming technology

 

  [1]    Pankaj K Mandal and Derrick J Rossi. (2013) Reprogramming human fibroblasts to pluripotency 

       using modified mRNA. Nature Protocols, 8: 568 – 582.

  [2]     Angel, M., and Yanik, M.F. (2010) Innate immune suppression enables frequent transfection with 

       RNA encoding reprogramming proteins. PLoS One 5: e11756.

  [3]     Warren, L., Manos, P.D., Ahfeldt, T., Loh, Y.H., Li, H., Lau, F., Ebina, W., Mandal, P.K., Smith, Z.D.,

        Meissner, A., Daley, D.Q., Brack, A.S., Collins, J.J., Cowan, C., Schlaeger, T.M., and Rossi, D.J. (2010)

       Highly efficient reprogramming to pluripotency and directed differentiation of human cells with 

       synthetic modified mRNA. Cell Stem Cell 7: 618-630.

  [4]     Yakubov, E., Rechavi, G., Rozenblatt, S., and Givol, D. (2010) Reprogramming of human fibroblasts

       to pluripotent stem cells using mRNA of four transcription factors. Biochem Biophys Res Commun.

      394: 189-193.


Additional Publications

●     Paull D; Sevilla A; Zhou H; Hahn AK; Kim H; Napolitano C; Tsankov A; Shang L; Krumholz K; 

 Jagaseesan P; Woodard CM; Sun B; Vilboux T; Zimmer M; Forero E; Moroziewicz DN; 

Martinez H; Malicdan MCV; Weiss KA; Vensand LB; Dusenberry CR; Polus H; Sy KTL; 

Kahler DJ; Gahl WA; Solomon SL; Chang S; Meissner A; Eggan K; Noggle SA. "Automated,

 high-throughput derivation, characterization and differentiation of induced pluripotent 

stem cells." Naure Methods 12:885 (2015)

●        Briggs SF; Dominguez AA; Chavez SL; Reijo Pera RA. "XIST in human preimplantation 

embryos and iPSCs." Stem Cells 33:1771 (2015)

 ●     Josowitz R, Lu J, Falce C, D’Souza SL, Wu M, Cohen N, Dubois NC; Zhao Y; Sobie EA; Fishman

 GI; Gelb BD. Identification and purification of human induced pluripotent stem cell-derived

 atrial-like cardiomyocytes based on sarcolipin expression. PLoS ONE 9(7): e101316 (2014)

 

产品列表
产品编号 产品名称 产品规格 产品等级 备注
00-0071 Stemgent® mRNA Reprogramming Kit
Stemgent® mRNA 重编程试剂盒		 1kit – –