StemRNA™ 3^rd Gen Reprogramming Kit

00-0076

Brand: Stemgent™

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The StemRNA 3^rd Gen Reprogramming Kit provides the fastest, most efficient method for generating clinically relevant iPS cells using a non-integrating, mRNA-based protocol. This technology supports generating iPSC lines derived from fibroblasts, blood, and urine using one multi-purpose kit.

Third Generation RNA Kit for Cellular Reprogramming of Fibroblasts, Blood, and Urine

The latest evolution in Stemgent RNA reprogramming, the StemRNA 3^rd Gen Reprogramming Kit (formerly called the StemRNA-NM Reprogramming Kit), combines non-modified RNA and microRNA technology to provide a kit for stem cell researchers with a new level of versatility, simplicity and time savings, enabling cellular reprogramming of human fibroblasts, and cells from blood and now urine for difficult to reprogram patient samples.

Key Benefits

• Flexible technology generates high-quality human iPS cell lines from multiple target cell types
  Out-of-the-box reprogramming of cells from skin (fibroblasts), blood (endothelial progenitor cells; EPCs) and urine (urine-derived progenitor cells; UPCs)
• High efficiency, non-integrating reprogramming
  StemRNA-3^rd Gen requires a few as four additional reagents. Double stranded microRNAs enhance reprogramming efficiency, providing iPS cells with high efficiency (up to 4% from fibroblasts, up to 3% from EPCs, up to 0.5% from UPCs).
• Time-saving protocol delivers faster results facilitating higher throughput
  Colonies ready to pick in 10-14 days from fibroblasts and 12-16 days from EPCs or UPCs. No screening needed

Timeline for StemRNA 3^rd reprogramming of fibroblasts

Download our StemRNA-3^rd Gen Reprogramming Posters:

ISSCR 2015

ISSCR 2016

Specifications

Product Name: StemRNA 3rd Gen Reprogramming Kit

Catalog Number: 00-0076

Storage and Stability: Store all three kit components at or below −70 °C. Kit components are stable for a minimum of 3 months from date of receipt when stored as directed.

Quality Control: The individual mRNAs are tested for size and integrity. The StemRNA 3^rd Gen Reprogramming Kit is functionally validated for successful RNA-based reprogramming of adult fibroblasts, human umbilical vein endothelial cells (HUVECs), EPCs, and UPCs. Complete reprogramming of iPS cell colonies is confirmed by expression of pluripotency markers and appropriate colony morphology. All components of the kit are sterile and have tested negative for Mycoplasma spp.

Recommended Usage: For use with associated StemRNA 3^rd Gen Reprogramming Kit protocols for generation of iPS cells.

Kit Contents:

• OKSMNL NM-RNA (Part No. 05-0040), 30 µg, 1 vial
• EKB NM-RNA (Part No. 05-0041), 22 µg, 1 vial
• NM-microRNAs (Part No. 05-0042), 15 µg, 1 vial

Documents

Protocols:

• Protocol: Stemgent StemRNA 3^rd Gen Reprogramming Kit for Reprogramming Adult and Neonatal Human Fibroblasts

• Protocol: Stemgent StemRNA 3^rd Gen Reprogramming Kit for Reprogramming Blood-Derived EPCs

• Protocol: Stemgent StemRNA 3^rd Gen Reprogramming Kit for Reprogramming Urine-Derived Progenitor Cells (UPCs)

Posters:

• ISSCR2015-StemRNA-NM-mobile.pdf

• ISSCR2016-StemRNA-NM_poster.pdf

FAQ:

• Frequently Asked Questions: REPROCELL Custom iPSC Services (FAQ-DISCOVERY-IPSCSERVICES-D001001.pdf)

References

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

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 3^rd 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 the elimination of reprogramming vectors.

Additional Publications

• Fiacco E, Alowaysi M; Astro V; Adamo A. Generation of an iPSC cohort of isogenic iPSC lines (46-XY and 47-XXY) from a non-mosaic Klinefelter Syndrome patient (47-XXY) (KAUSTi008-A, KAUSTi008-B, KAUSTi008-C, KAUSTi008-D, KAUSTi008-E, KAUSTi008-F, KAUSTi008-G). Stem Cell Res 50:102119 (2021).
• Fiacco E; Alowaysi M; Astro V; Adamo A. Derivation of two naturally isogenic iPSC lines (KAUSTi006-A and KAUSTi006-B) from a mosaic Klinefelter Syndrome patient (47-XXY/46-XY). Stem Cell Res 49:102049 (2020).
• Alowaysi M; Fiaccdo E; Astro V; Adamo A. Establishment of an iPSC cohort from three unrelated 47-XXY Klinefelter Syndrome patients (KAUSTi007-A, KAUSTi007-B, KAUSTi009-A, KAUSTi009-B, KAUSTi010-A, KAUSTi010-B). Stem Cell Res 49:102042 (2020).
• Isono W; Kawasaki T; Ichida JK; Ayabe T; hiraike O; Umezawa A; Akutsu H. The combination of dibenzazepine and a DOT1L inhibitor enables a stable maintenance of human naïve-state pluripotency in non-hypoxic conditions. Regen Therap 15:161-168 (2020).
• Alowaysi M; Fiacco E; Astro V; Adamo A. Establishment of iPSC lines from a high-grade Klinefelter Syndrome patient (49-XXXXY) and two genetically matched healthy relatives (KAUSTi003-A, KAUSTi004-A, KAUSTi004-B, KAUSTi005-A, KAUSTi005-B, KAUSTi005-C). Stem Cell Res 49:102008 (2020).
• Zorzan I; Pellergrini M; Arboit M; Incarnato I; Maldotti M; Forcato M; Malagoli Tagliazucci G; Carbognin E; Montagner M; Oliciero S; Martello G. The transcriptional regulator ZNF398 mediates pluripotency and epithelial character downstream of TGF-beta in human PSCs. Nature Commun 11:2364 (2020).
• Grosch M; Ittermann S; Rusha E; Greisle T; Ori C; Truong D-JJ; O'Niell AC; Pertek A; Westmeyer GG; Drukker M. Nucleus size and DNA accessibility are linked to the regulation of paraspeckle formation in cellular differentiation. BMC Biology 18:42 (2020).
• Wamaitha SE; Grybel KJ; Alanis-Lobato G; Gerri C; Ogushi S; McCarthy A; Kahdevaiah SK; Healy L; Lea RA; Molina-Arcas M; Devito LG; Elder K; Snell P; Christie L; Downward J; Turner JMA; Naikan KK. IGF1-mediated human embryonic stem cell self-renewal recapitulates the embryonic niche. Nature Commun 11:764 (2020).
• Gong J; Cai H; NYSCF Global Stem Array Team; Noggle S; Paull D; Rizzolo LJ; Del Priore LV; Fields MA. Stem cell-derived retinal pigment epithelium from patients with age-related macular degeneration exhibit reduced metabolism and matrix interactions. Stem Cells Transl Med :https://doi.org/10.1002/sctm.19-0321 (2019).
• Liu G; David BT; Trawczynski M; Fessler RG. Advances in pluripoptent stem cells: History, mechanisms, technologies, and applications. Stem Cell Rev Rep :doi.org/10.1007/s12015-019-09935-x (2019).
• Bredenkamp N; Yang J; Clarke J; Stirparo GG; von Meyenn F; Baker D; Drummond R; Ren Y; Li D; Wu C; Rostovskaya M; Eminli-Meissner S; Smith A; Guo G. Wnt Inhibition Facilitates RNA-Mediated Reprogramming of Human Somatic Cells to Naive Pluripotency. Stem Cell Rep 13:1083 (2019).
• 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).
• 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).
• 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).
• 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).
• 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).
• 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).
• 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).
• 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)
• 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)