Long complex steps, poor efficiency, expensive reagents, and scant results…don’t you wish that your cell-based experiments were a “little” bit easier and a “little” more efficient? These desires are commonplace for researchers generating induced pluripotent stem cells (iPSCs) in the hope of constructing patient-specific therapies. So what can we do to improve this important process?
In a recent Nature Methods paper, researchers from the laboratory of Nicola Elvassore (University of Padova, Italy) have described their “huge” step towards improving iPSC generation: scaling the entire process down to the microliter scale by employing a fabricated microfluidics chip platform.
Amazingly, they demonstrate that their automatable strategy can generate iPSCs at a level 50-fold better than traditional reprogramming strategies at a fraction of the monetary and labor costs!
So how did they accomplish this micro-marvel?
- The group fabricated a microfluidics chip containing multiple microliter-scale reaction chambers using multilayer soft lithography with a silicon-based organic polymer, which:
- Reduces the reaction volume from a typical 2 milliliters down to just 20 microliters
- Concentrates important growth factors released by cells during the reprogramming process to promote conversion of cells into the pluripotent state
- The microfluidic reprogramming process used the transient expression of modified mRNA of pluripotency-associated factors in human fibroblasts under chemically-defined, xeno- and feeder-free conditions
- While traditional mRNA reprogramming at the milliliter scale is inefficient and expensive, using the microfluidic chip led to 80% transfection rates and a 100-fold reduction in reagent requirements/costs
- Amazingly, reprogramming in the microfluidic chip boosted reprogramming efficiency from ~2 colonies per 100 starting cells using traditional techniques to ~120 pure and homogenous iPSCs colonies per 100 starting cells! Wow!
- Handily, microfluidic media processing can be safely automated and remotely controlled
- iPSCs could also rapidly differentiate into high-quality heart muscle cells and liver cells making the microfluidic chip highly relevant for future clinical application of iPSCs and their derivatives
Combine 36 microfluidics chambers onto a single chip whose functions can be remotely controlled and you get yourself a cheap and reliable means to perform (nearly) endless numbers of parallel reactions. The authors note that other than producing vast amounts of clinical-grade high-quality iPSCs and their derivatives, this system can assess the consequences of high precision microenvironment manipulation and the mechanisms behind the reprogramming process.
Here at Epigenie, we have previously shown how microfluidics can help chromatin immunoprecipitation and quantitative PCR, and this help now extends to the generation of iPSCs. So, always remember that a little can go a long way and check out microfluidics new trick at Nature Methods, April 2016.