ArticleStem cell research & therapy2013
Efficient expansion of human keratinocyte stem/progenitor cells carrying a transgene with lentiviral vector.
Article in Stem cell research & therapy, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 18 citations in OpenAlex.
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- Hypoxia and Foxn1 alter the proteomic signature of dermal fibroblasts to redirect scarless wound healing to scar-forming skin wound healing in Foxn1BMC biology · 2024Article
- Human Hematopoietic Stem Cells Co-cultured in 3D with Stromal Support to Optimize Lentiviral Vector-mediated Gene Transduction.Indian journal of hematology & blood transfusion : an official journal of Indian Society of Hematology and Blood Transfusion · 2023Article
- EGFR-mediated epidermal stem cell motility drives skin regeneration through COL17A1 proteolysis.The Journal of cell biology · 2021Article
- A new chemical complex can rapidly concentrate lentivirus and significantly enhance gene transduction.Cytotechnology · 2018Article
- P38 MAPK inhibition prevents polybrene-induced senescence of human mesenchymal stem cells during viral transduction.PloS one · 2018Article
- Generation of Genetically Modified Organotypic Skin Cultures Using Devitalized Human Dermis.Journal of visualized experiments : JoVE · 2015Article
- Feeder Layer Cell Actions and Applications.Tissue engineering. Part B, Reviews · 2015Review
- Cell motion predicts human epidermal stemness.The Journal of cell biology · 2015Article
- Long-term skin regeneration from a gene-targeted human epidermal stem cell clone.Molecular therapy : the journal of the American Society of Gene Therapy · 2014Article
- Optimization of lentiviral transduction parameters and its application for CRISPR-based secretome modification of human endometrial mesenchymal stem cells.Cell cycle (Georgetown, Tex.)Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
introductionThe development of an appropriate procedure for lentiviral gene transduction into keratinocyte stem cells is crucial for stem cell biology and regenerative medicine for genetic disorders of the skin. However, there is little information available on the efficiency of lentiviral transduction into human keratinocyte stem/progenitor cells and the effects of gene transduction procedures on growth potential of the stem cells by systematic assessment.
methodsIn this study, we explored the conditions for efficient expansion of human keratinocyte stem/progenitor cells carrying a transgene with a lentiviral vector, by using the culture of keratinocytes on a feeder layer of 3 T3 mouse fibroblasts. The gene transduction and expansion of keratinocytes carrying a transgene were analyzed by Western blotting, quantitative PCR, and flow cytometry.
resultsPolybrene (hexadiamine bromide) markedly enhanced the efficiency of lentiviral gene transduction, but negatively affected the maintenance of the keratinocyte stem/progenitor cells at a concentration higher than 5 μg/ml. Rho-assiciated kinase (ROCK) inhibitor Y-27632, a small molecule which enhanced keratinocyte proliferation, significantly interfered with the lentiviral transduction into cultured human keratinocytes. However, a suitable combination of polybrene and Y-27632 effectively expanded keratinocytes carrying a transgene.
conclusionsThis study provides information for effective expansion of cultured human keratinocyte stem/progenitor cells carrying a transgene. This point is particularly significant for the application of genetically modified keratinocyte stem/progenitor stem cells in regenerative medicine.
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