DEAE-Dextran Enhances the Lentiviral Transduction of Primary Human Mesenchymal Stromal Cells from All Major Tissue Sources Without Affecting Their Proliferation and Phenotype
-
Published:2022-08-23
Issue:
Volume:
Page:
-
ISSN:1073-6085
-
Container-title:Molecular Biotechnology
-
language:en
-
Short-container-title:Mol Biotechnol
Author:
Amadeo FrancescoORCID, Hanson Vivien, Murray PatriciaORCID, Taylor ArthurORCID
Abstract
AbstractGenetic engineering of mesenchymal stromal cells (MSCs) is a tool widely used to explore MSC properties in vitro and in vivo. Lentiviral infection with the use of polycations as an adjuvant is a method that is commonly used to generate stably transduced cells. However, it is known that some polycations can negatively affect primary MSCs and to date, no study has explored the effect of different polycations on the transduction efficiency and properties of all main types of MSCs, namely those derived from umbilical cord, bone marrow and adipose tissue. Here we explore a range of polycations, using transduction protocols with and without spinoculation, to produce stably transduced MSCs from these three tissue sources. We identified that an overnight incubation with diethylaminoethyl-dextran (DEAE-Dextran) is the protocol associated with the best transduction efficiency without compromising the viability of the cells, and which worked consistently with lentiviral particles encoding for different transgenes. Transduced and sorted MSC populations revealed no significant changes in proliferation, morphology and expression of MSC markers compared to naïve MSCs. Following this study, we conclude that DEAE-Dextran is a polycation that can be successfully used to enhance the transduction of MSCs from all major tissue sources.
Funder
H2020 Marie Skłodowska-Curie Actions
Publisher
Springer Science and Business Media LLC
Subject
Molecular Biology,Applied Microbiology and Biotechnology,Biochemistry,Bioengineering,Biotechnology
Reference25 articles.
1. Fung, M., Yuan, Y., Atkins, H., Shi, Q., & Bubela, T. (2017). Responsible translation of stem cell research: An assessment of clinical trial registration and publications. Stem Cell Reports, 8(5), 1190–1201. 2. Amadeo, F., Trivino Cepeda, K., Littlewood, J., Wilm, B., Taylor, A., & Murray, P. (2021). Mesenchymal stromal cells: What have we learned so far about their therapeutic potential and mechanisms of action? Emerging Topics in Life Sciences, 5(4), 549–562. 3. Niess, H., Thomas, M. N., Schiergens, T. S., Kleespies, A., Jauch, K. W., Bruns, C., Werner, J., Nelson, P. J., & Angele, M. K. (2016). Genetic engineering of mesenchymal stromal cells for cancer therapy: Turning partners in crime into Trojan horses. Innovative Surgical Sciences, 1(1), 19–32. 4. Scarfe, L., Taylor, A., Sharkey, J., Harwood, R., Barrow, M., Comenge, J., Beeken, L., Astley, C., Santeramo, I., Hutchinson, C., Ressel, L., Smythe, J., Austin, E., Levy, R., Rosseinsky, M. J., Adams, D. J., Poptani, H., Park, B. K., Murray, P., & Wilm, B. (2018). Non-invasive imaging reveals conditions that impact distribution and persistence of cells after in vivo administration. Stem Cell Research & Therapy, 9(1), 332. 5. Amadeo, F., Plagge, A., Chacko, A., Wilm, B., Hanson, V., Liptrott, N., Murray, P., & Taylor, A. (2022). Firefly luciferase offers superior performance to AkaLuc for tracking the fate of administered cell therapies. European Journal of Nuclear Medicine and Molecular Imaging, 49(3), 796–808.
Cited by
10 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献
|
|