RT Journal Article
SR Electronic
T1 Micropatterned substrates to promote and dissect reprogramming of human somatic cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 111369
DO 10.1101/111369
A1 Jared Carlson-Stevermer
A1 Ty Harkness
A1 Ryan Prestil
A1 Stephanie Seymour
A1 Gavin Knight
A1 Randolph Ashton
A1 Krishanu Saha
YR 2017
UL http://biorxiv.org/content/early/2017/05/08/111369.abstract
AB Reprogramming of human somatic cells to induce pluripotent stem cells (iPSCs) generates valuable precursors for disease modeling and regenerative medicine. However, the reprogramming process can be inefficient and noisy, creating many partially reprogrammed cells in addition to fully reprogrammed iPSCs. To address these shortcomings, we developed a micropatterned substrate that allows for dynamic live-cell microscopy of thousands of cell subpopulations undergoing reprogramming. Micropatterning facilitated a change in shape, size and clustering of nuclei to promote somatic identity erasure. Increased proliferation, cell density and decreased intercellular YAP signaling accompanied these nuclear changes. A combination of eight nuclear characteristics could be used to track reprogramming progression and distinguish partially reprogrammed cells from those that were fully reprogrammed.Micropatterned substrates constitute a new tool for facile iPSC production and can be used in high-throughput to probe and understand the subcellular changes that accompany human cell fate transitions.