ArticleJournal of visualized experiments : JoVE2011
Transfecting and nucleofecting human induced pluripotent stem cells.
Article in Journal of visualized experiments : JoVE, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 26 citations in OpenAlex.
- Improving viability and transfection efficiency with human umbilical cord wharton's jelly cells through use of a ROCK inhibitor.Cellular reprogramming · 2014Trial
- Detecting and measuring of GPCR signaling - comparison of human induced pluripotent stem cells and immortal cell lines.Frontiers in endocrinology · 2023Review
- Protocol for a Wnt reporter assay to measure its activity in human neural stem cells derived from induced pluripotent stem cells.Current research in neurobiology · 2023Article
- Targeted mutagenesis in human iPSCs using CRISPR genome-editing tools.Methods (San Diego, Calif.) · 2021Article
- Transgene Delivery to Human Induced Pluripotent Stem Cells Using Nanoparticles.Pharmaceuticals (Basel, Switzerland) · 2021Review
- Video-based calcification assay: A novel method for kinetic analysis of osteogenesis in live cultures.MethodsX · 2021Article
- Transfection of hPSC-Cardiomyocytes Using Viafect™ Transfection Reagent.Methods and protocols · 2020Article
- Integration of nano- and biotechnology for beta-cell and islet transplantation in type-1 diabetes treatment.Cell proliferation · 2020Review
- Disease-associated mutations of claudin-19 disrupt retinal neurogenesis and visual function.Communications biology · 2019Article
- Low Osteogenic Yield in Human Pluripotent Stem Cells Associates with Differential Neural Crest Promoter Methylation.Stem cells (Dayton, Ohio) · 2018Article
- Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles.International journal of nanomedicine · 2018Article
- Biological Effects of Culture Substrates on Human Pluripotent Stem Cells.Stem cells international · 2016Review
- Applications of stem cells in interdisciplinary dentistry and beyond: an overview.Annals of medical and health sciences research · 2013Article
- An in vitro mechanism study on the proliferation and pluripotency of human embryonic stems cells in response to magnesium degradation.PloS one · 2013Article
- Derivation of neural progenitors and retinal pigment epithelium from common marmoset and human pluripotent stem cells.Stem cells international · 2012Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genetic modification is continuing to be an essential tool in studying stem cell biology and in setting forth potential clinical applications of human embryonic stem cells (HESCs). While improvements in several gene delivery methods have been described, transfection remains a capricious process for HESCs, and has not yet been reported in human induced pluripotent stem cells (iPSCs). In this video, we demonstrate how our lab routinely transfects and nucleofects human iPSCs using plasmid with an enhanced green fluorescence protein (eGFP) reporter. Human iPSCs are adapted and maintained as feeder-free cultures to eliminate the possibility of feeder cell transfection and to allow efficient selection of stable transgenic iPSC clones following transfection. For nucleofection, human iPSCs are pre-treated with ROCK inhibitor, trypsinized into small clumps of cells, nucleofected and replated on feeders in feeder cell-conditioned medium to enhance cell recovery. Transgene-expressing human iPSCs can be obtained after 6 hours. Antibiotic selection is applied after 24 hours and stable transgenic lines appear within 1 week. Our protocol is robust and reproducible for human iPSC lines without altering pluripotency of these cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.