Reduce Hands-on Time with the CytoTune®-iPS Sendai Reprogramming Kit
Click on the tabs below to find out how you can experience successful reprogramming in one simple transduction. |
Comparison of reprogramming workflows using the CytoTune®-iPS Sendai Kit, mRNA, and lentivirus
Generation of iPSCs in 3-4 weeks is accomplished after a single transduction with the CytoTune® kit, while mRNA requires 17 sequential days of transduction. Traditional reprogramming methods such as with lentivirus or retrovirus can require multiple transductions for iPSC generation.
Protocol timeline for iPSC generation from human primary fibroblasts with the CytoTune®-iPS Sendai Reprogramming Kit
A single transduction with the CytoTune® kit generates iPSC colonies typically 3-4 weeks post-transduction. For a detailed protocol, please consult the product handbook.
| Day -2 | Plate fibroblasts in fibroblast culture medium (DMEM + 10% FBS) into two wells of a 6-well plate to achieve 5 x 105 cells/well on day 3. |
| Day 0 | Add CytoTune®-iPS Reprogramming Kit contents to your cells. |
| Day 1-7 | Replace the fibroblast culture medium (DMEM + 10% FBS) with fresh fibroblast culture medium on day 4, and then every other day thereafter. |
| Day 7 | Harvest cells with trypsin, then plate the cells onto MEF feeder-coated plates in fibroblast culture medium. |
| Day 8 | Replace the fibroblast medium with complete KnockOut™ Serum Replacement medium. |
| Day 9-25 | Observe the cells and replace the spent medium daily. Look for the emergence of cell clumps indicative of iPSCs. |
| 3-4 weeks post-transduction | Perform live staining with Tra1-60 or Tra1-81 to select reprogrammed colonies. Manually pick colonies and transfer them to fresh MEF plates. |
Simple generation of iPSC colonies from human neonatal foreskin fibroblast cells (strain BJ) with the CytoTune®-iPS Sendai Reprogramming Kit
(A) Fibroblast cells prior to transduction. (B-D) iPSC colonies at (B) 3 weeks and (C) 4 weeks post-transduction with a single application of the CytoTune®-iPS Sendai Reprogramming Kit can be easily identified due to the high efficiency and low background of partially reprogrammed cells. (D) iPSC colony after manual picking and transfer to KnockOut™ SR Medium.
Learn more about the CytoTune® kit
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For research use only. Not for human or animal therapeutic or diagnostic use.
