High-efficiency cellular reprogramming with microfluidics.
Camilla LuniStefano GiulittiElena SerenaLuca FerrariAlessandro ZambonOnelia GaglianoGiovanni G GiobbeFederica MichielinSebastian KnöbelAndreas BosioNicola ElvassorePublished in: Nature methods (2016)
We report that the efficiency of reprogramming human somatic cells to induced pluripotent stem cells (hiPSCs) can be dramatically improved in a microfluidic environment. Microliter-volume confinement resulted in a 50-fold increase in efficiency over traditional reprogramming by delivery of synthetic mRNAs encoding transcription factors. In these small volumes, extracellular components of the TGF-β and other signaling pathways exhibited temporal regulation that appears critical to acquisition of pluripotency. The high quality and purity of the resulting hiPSCs (μ-hiPSCs) allowed direct differentiation into functional hepatocyte- and cardiomyocyte-like cells in the same platform without additional expansion.
Keyphrases
- induced pluripotent stem cells
- high efficiency
- induced apoptosis
- transcription factor
- signaling pathway
- high throughput
- endothelial cells
- cell cycle arrest
- pi k akt
- transforming growth factor
- endoplasmic reticulum stress
- angiotensin ii
- oxidative stress
- circulating tumor cells
- gene expression
- liver injury
- copy number
- cell proliferation
- genome wide
- embryonic stem cells
- drug induced