Micropatterned substrates to promote and dissect reprogramming of human somatic cells

Abstract

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.

Targeting cell plasticity for regeneration: From in vitro to in vivo reprogramming

“…2C). Similarly, Saha and colleagues demonstrated a micropatterning based method to dissect and understand the intermediate cell populations as well as the dynamics of the nuclear reprogramming process [124]. This study used micropatterns to modulate nuclear size and shape and established a set of nuclear characteristics to distinguish between fully and partially reprogrammed cells.…”