Stemgent^® mRNA 重编程试剂盒

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

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)