High-efficiency cellular reprogramming with microfluidics
@article{Luni2016HighefficiencyCR, title={High-efficiency cellular reprogramming with microfluidics}, author={Camilla Luni and Stefano Giulitti and Elena Serena and Luca Ferrari and Alessandro Zambon and Onelia Gagliano and Giovanni Giuseppe Giobbe and Federica Michielin and S Kn{\"o}bel and Andreas Bosio and Nicola Elvassore}, journal={Nature Methods}, year={2016}, volume={13}, pages={446-452} }
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…
82 Citations
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References
SHOWING 1-10 OF 37 REFERENCES
Functional differentiation of human pluripotent stem cells on a chip
- Biology, EngineeringNature Methods
- 2015
A robust microfluidics-based approach is described for the differentiation of human pluripotent stem cells directly on a chip for functional phenotypes and responses to temporally defined drug treatments.
Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA.
- BiologyCell stem cell
- 2010
How microRNAs facilitate reprogramming to pluripotency
- BiologyJournal of Cell Science
- 2012
Recent findings that show that, in addition to transcription factors, microRNAs can promote pluripotent reprogramming and can even substitute for these pluripotency transcription factors in some cases are discussed.
A comparison of non-integrating reprogramming methods
- BiologyNature Biotechnology
- 2015
Comparing Sendai-viral (SeV), episomal (Epi) and mRNA transfection mRNA methods for generating integration-free hiPSCs provides a valuable resource to inform the use of specific reprogramming methods for different laboratories and different applications, including clinical translation.
Small molecules facilitate rapid and synchronous iPSC generation
- BiologyNature Methods
- 2014
This work shows that exposure of OKSM-expressing cells to both ascorbic acid and a GSK3-β inhibitor (AGi) facilitates more synchronous and rapid iPSC formation from several mouse cell types, enabling dissection of the underlying mechanisms in more homogeneous cell populations.
Adhesive Signature-based, Label-free Isolation of Human Pluripotent Stem Cells
- BiologyNature Methods
- 2013
We demonstrate substantial differences in 'adhesive signature' between human pluripotent stem cells (hPSCs), partially reprogrammed cells, somatic cells and hPSC-derived differentiated progeny. We…
Combinatorial Modulation of Signaling Pathways Reveals Cell-Type-Specific Requirements for Highly Efficient and Synchronous iPSC Reprogramming
- BiologyStem cell reports
- 2014
Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA
- BiologyScientific reports
- 2012
This work accelerated the mRNA reprogramming process through stepwise optimization of the RF cocktail and leveraged these kinetic gains to establish a feeder-free, xeno-free protocol which slashes the time, cost and effort involved in iPSC derivation.
Microfluidic Control of Cell Pairing and Fusion
- BiologyNature Methods
- 2009
A microfluidic device to trap and properly pair thousands of cells, and observed that electrical fusion was more efficient than chemical fusion, with membrane reorganization efficiencies of up to 89%.