StemRNA™ 3rd Gen Reprogramming Kit
第三代 RNA 重新編程試劑試劑盒
可用於皮膚的成纖維細胞 (fibroblasts)﹑血液和尿液，共 3 種細胞，高效率生成 iPS 細胞。重新編程時間短，在 10-14 天之內從成纖維細胞中挑選出菌落，EPC 或 UPC 需 12-16 天。無需篩選。，無外來基因影響，無病毒殘留，無需餵養細胞，針對纖維細胞有完整 xeno-free 工作流程。
分類：mRNA Reprogram > 幹細胞相關產品 > 試劑溶液
標籤：iPS, Reprogramming, 幹細胞, 重編程
試劑特色
- 市面上最經濟的選擇
- 高效率生成 iPS 細胞
- 重新編程時間短
- 無外來基因影響
- 無病毒殘留
- 無需餵養細胞
- 針對纖維細胞有完整 xeno-free 工作流程
三種細胞來源比較
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| StemRNA-3rd Gen Reprogramming Kit (00-0076) | |||
|---|---|---|---|
| Feature | Fibroblasts | Urine (UPCs) | Blood (EPCs) |
| No. wells per kit | 9 | 3 | 3 |
| No Transfections required | 4 | 6-8 | 6-8 |
| Days to primary iPS cell colonies | 10-14 | 12-14 | 12-14 |
| Reprogramming efficiency | 2~4% | 0.1~0.5% | 0.4~3% |
| Screening Required | No | No | No |
| Xeno-compatible protocol | Yes | No | No |
實驗時間軸
(以纖維細胞 fibroblast 為例)
優化過的實驗步驟，可以在 10~12 天獲得 iPS 細胞
iPS 細胞型態
(以纖維細胞 fibroblast 為例)
建立的 iPS 細胞，繼代 7 次之後，依然呈現良好的細胞型態。
iMatrix 511 iPS
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| 分類 | 內容 | 備註 |
|---|---|---|
| 細胞種類 | 人類纖維細胞 | Human fibroblasts |
| 使用試劑 | StemRNA NM reprogramming kit | ReproCELL cat. # 00-0076 |
| 培養溶液 | NutriStem™ XF/FF medium | ReproCELL cat. # 01-0005 |
| 塗布試劑 | iMatrix-511 | ReproCELL cat. # NP392-011 |
初代 iPS 細胞型態
分別從皮膚、血液、尿液分離出的細胞，以 StemRNA™ 3rd Gen Reprogramming Kit 進行重新編程。可看到初代 iPS 細胞株的型態完整，並以幹細胞特定抗體染色標定確認。(StainAlive™ TRA-1-60 Antibody (ReproCELL cat. # 09-0068).)
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| 貨號 | 內容物 |
|---|---|
| 00-0076 | StemRNA™ 3rd Gen Reprogramming Kit
|
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 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
- Shimada M; Tsukada K; Kagawa N; Matsumoto Y. Reprogramming and differentiation-dependent transcriptional alteration of DNA damage response and apoptosis genes in human induced pluripotent stem cells. J Radiation Res 2019:1-10 (2019).
- Bredenkamp N; Yang J; Clarke J; Stirparo GG; von Meyenn F; Baker D; Drummond R; Li D; Wu C; Rostovskaya M ; Smith A; Guo G. Efficient RNA-mediated reprogramming of human somatic cells to naïve pluripotency facilitated by tankyrase inhibition. bioRxiv https://doi.orb/10.1101/636670: (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)
Specification Sheets
Safety Data Sheets
Protocols
- Protocol: Stemgent StemRNA 3rd Gen Reprogramming Kit for Reprogramming Adult and Neonatal Human Fibroblasts
- Protocol: Stemgent StemRNA-3rd Gen Reprogramming Kit for Reprogramming Blood-Derived EPCs
- Protocol: Stemgent StemRNA 3rd Gen Reprogramming Kit for Reprogramming Urine-Derived Progenitor Cells (UPCs)
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