High-Efficiency Cellular Reprogramming by Nanoscale Puncturing

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journal

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Detail(s)

Original languageEnglish
Pages (from-to)5473-5481
Journal / PublicationNano Letters
Volume20
Issue number7
Online published10 Jun 2020
Publication statusPublished - 8 Jul 2020

Abstract

Induced pluripotent stem cells (iPSCs) bear great potential for disease modeling, drug discovery, and regenerative medicine; however, the wide adoption of iPSC for clinically relevant applications has been hindered by the extremely low reprogramming efficiency. Here, we describe a high-efficiency cellular reprogramming strategy by puncturing cells with an array of diamond nanoneedles, which is applied to temporally disrupt the cell membrane in a reversible and minimally invasive format. This method enables high-efficiency cytoplasmic delivery of mini-intronic plasmid vectors to initiate the conversion of human fibroblast cells to either primed iPSCs or naıve iPSCs. The nanopuncturing operation is directly performed on cells in ̈ adherent culture without any cell lift-off and is completed within just 5 min. The treated cells are then cultured in feeder-free medium to achieve a reprogramming efficiency of 1.17 ± 0.28%, which is more than 2 orders of magnitude higher than the typical results from common methods involving plasmid delivery.

Research Area(s)

  • cell reprogramming, induced pluripotency, intracellular delivery, diamond nanoneedle, membrane disruption