“More for less” is a phrase that generally gets most people excited, and scientists trying to make induced pluripotent stem cells (iPSCs) are no exception. The reprogramming of somatic cells to generate iPSCs aims to bring patient-specific cell replacement therapies to the masses, but the process currently suffers from obstacles common to new technologies – slow pace, low efficiency, high costs, and back breaking effort.
However, a recent report from the laboratory Guokai Chen in Scientific Reports has described a new reprogramming protocol which promises cheap, quick, and easy iPSC generation for all.
The improvements described in this new study include:
- Spin transduction-mediated reprogramming using the CytoTune™-iPS 2.0 Sendai RNA virus Reprogramming Kit (Life Technologies)
- Utilizes three different non-integrative vectors (polycistronic Klf4–Oct4–Sox2, cMyc, and Klf4)
- One kit can reprogram 24-48 samples generating 600 iPSC colonies per sample
- Low reagent cytotoxicity
- Small number of input cells (1 x 104) required
- A streamlined reprogramming schedule, which allows the initiation and synchronization of 20 simultaneous reprogramming experiments
- Morphological changes at day 2 allows the evaluation of transduction success
- Expansion of colonies using a serial dilution method onto plates directly coated with Matrigel in chemically defined E8 xeno-free feeder-free medium (Life Technologies)
- Allows passaging of single colonies by EDTA/PBS with ROCK inhibitors to reduce time
- Generates high purity cells for expansion/preservation at passage two.
- Highly efficiency media usage to reduces costs further
- Provision for multiple backups of parental cells and iPSCs, allowing for quick experiment repetition
- Cryopreservation of iPSCs directly on culture plates, to reduce costs and time, and expand preservation capacity.
Overall, the group has found that their enhancements can:
- Generate hundreds of iPSCs lines in a short time period
- Decrease experimental scale by 50-fold
- Increase number of reprogramed samples by 10-fold
- Decrease medium usage by 70%
- Decreased reagent cost by more than 80% per sample
- Drastically reduce cell culture workload
The streamlining of this previously daunting process will hopefully bring this exciting technology to labs great and small across the globe. See how you can apply these techniques to your iPSC experiments today! – Sci Rep, June, 2015.