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Human induced pluripotent stem cells (hiPSCs) play a crucial role in regenerative medicine and disease modeling, but their efficacy hinges on the efficient and safe delivery of exogenous genetic material. Traditional viral and electroporation methods still have limitations, especially in terms of biosafety and regulatory barriers, poor transfection accuracy, and high cytotoxicity after transfection. This study introduces nanotube (NT)-mediated nanoinjection as an effective non-viral strategy for the delivery of functional messenger RNA (mRNA) to high pluripotent stem cells (hiPSCs). To achieve nanoinjection into high pluripotent stem cells (hiPSCs), the study implemented delayed extracellular matrix (ECM) application and advanced surface engineering, combined with a redesigned NT design, using a large cargo storage capacity to enhance loading capacity and sharp edge geometry to improve the cell interface. Using mCherry, GFP, and YPet mRNA (average transfection rate: approximately 55% to approximately 64%), the study effectively injected nanoparticles into high pluripotent stem cells, including co-transfection of mCherry and GFP mRNA (approximately 61%). Through the excellent pluripotency marker expression (NANOG, OCT4, SOX2) and neuronal differentiation ability after multiple passages, the integrity of hiPSCs after nanoinjection was confirmed. This proof-of-concept study demonstrates that nanoinjection is a powerful tool for delivering mRNA to high expression rate pluripotent stem cells and establishes nanoinjection as a platform for precise cell engineering and functional stem cell applications. This research was published under the title "Poking Pluripotency: Nanoinjection Into Human iPSCs" in Advanced Materials.
References:
DOI: 10.1002/adma.202521046
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