StemRNA NM Reprogramming Kit

Product Classification : RNA Reprogramming for hiPSCResearch Theme : ES/iPS cells

RNAリプログラミングヒトiPS細胞(線維芽細胞・血液・尿用)
00-0076 - Overview

Fast and easy iPS reprogramming from fibroblasts, blood derived cells, urine derived cells

Features

 ・ Best cost efficiency on the market
 ・ Significant reduction in labor and time requirements
 ・ GMP compatible
 ・ No genomic artifacts
 ・ Virus free
 ・ Feeder-free
・ Entirely xeno-free work-flow for fibroblasts


Overview

Optimized reprogramming protocols for different cell types ensure reliable iPSC establishment within 10-12 days

Fig. 1: Timeline for fibroblast StemRNA NM reprogramming

Day 0: Plate Fibroblasts in Fibroblast Expansion Medium.
Day 1: Media switch to NutriStem Medium.
Day 1-4: Daily NM-RNA cocktail overnight transfections.
Optional on Day 8-14: Identify emerging iPSC colonies by TRA-1-60 live stain.
Day 10-14: Pick primary Fibroblast-NM-RNA-iPSC colonies, replate in NutriStem Medium.


Fig. 2: Timeline for blood derived EPC StemRNA NM reprogramming

Day 0: Plate EPCs.
Day 1-8: Daily NM-RNA cocktail transfection overnight.
Day 10: Switch to NutriStem medium.
Optional: Identify emerging iPSC colonies by TRA-1-60 live stain
Day 12: Continue culture in NutriStem medium
Day 12-16: Pick primary EPC-NM-RNA-iPSC colonies, replate in NutriStem Medium.


Fig. 3: Timeline for urine derived cell StemRNA NM reprogramming

Day 0: Plate UDCs.
Day 1-8: Daily NM-RNA cocktail transfection overnight.
Day 10: Switch to NutriStem Medium.
Optional: Identify emerging iPSC colonies by TRA-1-60 live stain.
Day 12: Continue culture in NutriStem Medium.
Day 12-16: Pick primary UDC-NM-RNA-iPSC colonies, replating in NutriStem Medium.



Fig. 4: Healthy iPSC colony morphology 7 passages after establishment

StemRNA NM reprogrammed iPSC colony at passage 7

Human fibroblasts were reprogrammed, expanded and maintained using StemRNA NM reprogramming kit (ReproCELL cat. # 00-0076), NutriStem™ XF/FF medium (ReproCELL cat. # 01-0005) and iMatrix-511 (ReproCELL cat. # NP392-011). Reprogrammed iPSCs at passage 7 show typical healthy colony morphology with clearly defined borders.



Fig. 5: Primary iPSC colony morphology and pluripotency


Plating densities and primary iPSC colony morphology for human fibroblasts (Fig. 5.A), blood derived endothelial progenitor cells (EPCs; Fig. 5.B), and urine derived cells (Fig. 5.C). Cells were reprogrammed and expanded using StemRNA NM reprogramming kit (ReproCELL cat. # 00-0076), NutriStem™ XF/FF medium (ReproCELL cat. # 01-0005) and iMatrix-511 coating substrate (ReproCELL cat. # NP392-011). Primary iPSC colonies were stained with live pluripotency marker StainAlive™ TRA-1-60 Antibody (ReproCELL cat. # 09-0068).



StemRNA NM Reprogramming Features

Product and Order Info

Cat. # Product Content Price
00-0076

Stemgent StemRNA-NM Reprogramming Kit

Contents
  • OSKMNL NM-RNA
  • NM-microRNAs
  • EKB NM-RNA
Kit
3 Components
Please inquire
01-0005 NutriStem™ XF/FF 500 mL Please inquire
RCHEOT002
/ RCHEOT005
bFGF
/ bFGF Xeno-Free
25 μg
/ 25 μg
Please inquire
09-0068 StainAlive™ TRA-1-60 Antibody (DyLight™ 488), Mouse anti-Human 100 μL Please inquire
NP892-011 iMatrix-511 (350 μg) 350 μg (2×175 μg) Please inquire
Use of a hypoxic 5% O2 incubator is strongly recommended for maximum reprogramming efficiencies.

Other ReproCELL RNA Reprogramming Technologies

Products
microRNA enhanced mRNA Reprogramming
Fast and efficient virus-free, DNA-free fibroblast reprogramming.
Self-replicative RNA Reprogramming
Self-Replicative RNA mediated, reliable reprogramming of blood-derived cells.

Related Products

Products
NutriStem™ XF/FF
Xeno-free medium for feeder-free iPSC/ESC reprogramming, expansion, and maintenance.
iMatrix-511
Laminin-511E8 fragment coating offers xeno-free, ideal growth conditions for feeder-free culture of iPSC, ESC, and many other cell types.
Laminin-5
Xeno-free coating substrate for feeder-free hiPSC/hESC culture. Laminin-332 derived.
bFGF Xeno-Free(FGF-2)
Xeno-free, human recombinant, high purity bFGF (basic fibroblast growth factor) for hiPSC/hESC culture.
bFGF
Human recombinant, high purity bFGF (basic fibroblast growth factor) for hiPSC/hESC culture.
ReproCryo DMSO Free™
Cutting-edge DMSO-free, xeno-free slow-freeze freezing medium for hiPS/hES cells.

Publications

  • Poleganov et al (2015) Efficient Reprogramming of Human Fibroblasts and Blood-Derived Endothelial Progenitor Cells Using Non modified RNA for Reprogramming and Immune Evasion. Hum Gene Ther. 11:751-66.